Isolation and expression of rainbow trout (Oncorhynchus mykiss) ovarian cDNA encoding 3β-hydroxysteroid dehydrogenase/Δ5−4-isomerase

A distinct shift in steroidogenesis from testosterone to 17α-hydroxyprogesterone occurs in the salmonid ovarian thecal cell layers immediately prior to oocyte maturation, and is a prerequisite for the production of 17α,20β-dihydroxy-4-pregnen-3-one (maturation-inducing hormone of salmonid fishes) by granulosa cells during oocyte maturation. 17α-Hydroxylase/17,20-lyase cytochrome P-450 (P-450_17α) and 3β-hydroxysteroid dehydrogenase/Δ^5?4-isomerase (3β-HSD) are the two major steroidogenic enzymes involved in the production of 17α-hydroxyprogesterone and testosterone. Using mammalian cDNA probes, we isolated and characterized full-length cDNAs encoding these two enzymes from a rainbow trout (Oncorhynchus mykiss) ovarian thecal cell cDNA library. The cloning of 2.4-kilobase cDNA encoding P-450_17α and transient expression of this clone in nonsteroidogenic monkey kidney tumor COS-1 cells have recently been reported (Sakai et al. 1992). We have isolated a 1.4-kilobase cDNA which is hybridized to the mammalian 3β-HSD cDNAs. Expression of this cDNA in COS-1 cells led to the production of an enzyme which is capable of converting dehydroepiandrosterone to androstenedione. In this study, enzymatic activities and expression of rainbow trout ovarian P-450_17α and 3β-HSD are discussed in relation of the steroidogenic shift occurring in the ovarian follicle layers.     

Morpho-physiological predictors of ovulatory success in white sturgeon,Acipenser transmontanus Richardson

Body length, oocyte diameter, germinal vesicle position, in vitro oocyte maturation response to progesterone, and plasma concentrations of progesterone, cortocosteroids, and 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-diOHP) were examined in white sturgeon females prior to their spawning induction and correlated with their subsequent ovulatory response. The relationship between broodfish size and ovulatory success was insignificant. Responsive females had larger oocytes and elevated plasma concentrations of corticosteroids and 17α,20β-diOHP, but did not differ significantly from nonresponsive females in progesterone concentrations. Germinal vesicle position and in vitro oocyte maturation response exhibited the closest relationships with ovulation, and can be used as practical predictors of ovulation for hormonally induced spawning of white sturgeon.     

Preliminary Observations on the Reproductive Physiology of Female Orangemouth Corvina in Captivity

Seasonal changes In oocyte growth and plasma estradiol, testosterone, thyroid hormones and vitellogenin levels were monitored in three captive adult female orangemouth corvina Cynoscion xanthulus subjected to a condensed (8 mo) seasonal cycle of photoperiod and water temperature. Plasma concentrations of estradiol and testosterone began to rise when temperature increased to 28 C and photoperiod to 15 h light (midsummer conditions). This was accompanied by elevated circulating levels of vitellogenin and the appearance of vitellogenic oocytes (diameter > 100 μm). Estradiol concentrations and mean oocyte size increased concurrently during late summer conditions and were maximum during fall conditions, approximately 8 wk after the beginning of ovarian recrudescence. In contrast, plasma levels of thyroid hormones did not show any distinct seasonal changes. Gonadally recrudesced females contained several size classes of vitellogenic oocytes. Injection of these fish with a luteinizing hormone-releasing hormone analog (LHRHa) caused maturation and spawning of the largest oocytes (mean diameter of follicles × 2500 μm). Another group of vitellogenic oocytes had been recruited into this size class by 2 wk after the end of spawning, which suggests that this species is capable of repeated spawning during the reproductive season. Injection of LHRHa resulted in increased plasma levels of gonadotropin and repeated spawning over several days. LHRHa-induced final oocyte maturation, ovulation and spawning were preceded by increases in plasma levels of two teleostean maturation-inducing steroids, 17α,20β, 21-trihydroxy-4-pregnen-3-one (20β-S) and 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-P₄). The results provide preliminary evidence that oocyte growth is stimulated in orangemouth corvina subjected to an abbreviated seasonal cycle in captivity under midsummer photoperiod and temperature conditions and is associated with seasonal increases in plasma estradiol concentrations and vitellogenin production. However, the relative importance of 20β-S versus 17α, 20β-P₄ in the control of final oocyte maturation in this species could not be determined from the results of the present study.     

Brain steroid contents in the catfish Heteropneustes fossilis: sex and gonad stage-specific changes

Neurosteroids are those which are synthesized in the central nervous system independently of supply by peripheral endocrine glands. In the present study, brain contents of the steroid hormones, estradiol-17β (E(2)), testosterone (T), corticosteroids, and progestins were investigated in both male and female catfish Heteropneustes fossilis in prespawning (vitellogenic) and spawning (post-vitellogenic) phases using ELISA or HPLC. The data show that the measured steroid hormones showed both stage-specific and sex-related variations. Brain E(2) was significantly higher in males in the prespawning phase and in females in the spawning phase. Testosterone was significantly higher in males in comparison with females in the prespawning phase. Cortisol was significantly higher in the prespawning and spawning phases in males than in females. Corticosterone level was low in the brain. 21-deoxycortisol and deoxycorticosterone were significantly higher in the prespawning phase than in the spawning phase. Male brain recorded the highest concentration of deoxycorticosterone. Progesterone (P(4)) was high in the prespawning phase and low in the spawning phase in both sexes. Levels of 17-hydroxy-4-pregnene-3,20-dione and 17,20β-dihydroxy-4-pregnen-3-one (17,20β-DP) and the metabolites of P(4) were the highest in females in the prespawning phase. The stage-specific and sexual differences in the content of the steroids suggest their biosynthesis in the brain, which may have implications in brain functions, in addition to reproductive regulation.     

Diurnal rhythm of steroid biosynthesis in the testis of terminal phase male of protogynous wrasse, Pseudolabrus sieboldi, a daily spawner

The wrasse, Pseudolabrus sieboldi, is a diandric protogynous labrid fish. Spawning is performed by a terminal phase (TP) male and an initial phase (IP) female between 6:00 and 9:00 h daily during two-month-long spawning season. In the present study, to investigate the roles of steroid hormones in the diurnal spermatogenesis of the P. sieboldi TP male, all steroid hormones produced in the testis were identified and the synthetic pathways of these steroids were determined. Furthermore, the circulating levels of the major steroids produced were analyzed throughout a day at 3-hour intervals during spawning season. In the testis, 11-ketotestosterone (11-KT), estradiol-17β (E2), 17,20β-dihydoxy-4-pregnane-3-one (17,20β-P) and 17,20β,21-trihydroxy-4-pregnen-3-one (20β-S) were synthesized as the major metabolites. In vitro steroid biosynthesis experiments showed similar results to the circulation profiles of the major steroids. This study is the first to clarify the complete steroidogenic pathways in the gonads of a diandric protogynous species throughout its life, when combined with the results of the steroidogenesis in the ovarian follicles. This is also the first report of a clear diurnal rhythm of the steroid production corresponding to the spermatogenic process in the testis of a male teleost.     

White sturgeon spawning and discharge augmentation

Abstract Observed spawning events of endangered Kootenai River white sturgeon, Acipenser transmontanus (Richardson), during 1994–2002 were examined to evaluate the effects of discharge from Libby Dam on spawning. Discharge from Libby Dam was manipulated in most years to provide enhanced flows (hereafter, augmented discharge) for white sturgeon spawning. The annual onset of spawning appeared independent of augmented discharge because initial spawning occurred prior to augmented discharge in 4 of 6 years. Spawning began 2–11 days after the highest river discharge in 4 of 9 years. Linear regression analysis indicated the onset of spawning was positively related to mean daily discharge from the dam but not mean daily temperature, 1‐day change in discharge (Δ discharge) or 3‐day Δ discharge. Logistic regression analysis suggested the probability of a spawning event was influenced by Julian date, mean daily water temperature, mean daily discharge and the 7‐day Δ discharge, but the predictive indices were small. Minor fluctuations in temperature and discharge characteristics had little additional predictive benefit for spawning after the river reached spawning thresholds. The highest probability of spawning (0.48) was for the temperature interval 9.5–9.9 °C, and 93% of the estimated spawning events occurred above 8 °C. Sixty percent of the estimated spawning events occurred at discharges ≥600 m³ s^?1, which comprised 45% of the range of discharge values.     

Molecular endocrinology of oocyte growth and maturation in fish

Pituitary gonadotropins (GTHs) are of primary importance in triggering oocyte growth and maturation. However, the actions of GTHs are not direct, but are mediated by the ovarian production of steroidal mediators of oocyte growth (estradiol-17β) and maturation (maturation-inducing hormone, MIH; 17α,20β-dihydroxy-4-pregnen-3-one, 17α,20β-DP in salmonid fishes; 17α,20β,21-trihydroxy-4-pregnen-3-one, 20β-S in sciaenid fishes). It is established that production of estradiol-17β and 17α,20β-DP by salmonid ovarian follicles occurs via the interaction of two cell layers, the thecal and granulosa cell layers (two-cell type model). A distinct shift in the salmonid steroidogenesis from estradiol-17β to 17α,20β-DP occurs in the ovarian follicle layer immediately prior to oocyte maturation. It is possible that this shift is a consequence of dramatic changes in the expression of the genes encoding various steroidogenic enzymes. As an initial step to address this question, we have isolated and characterized the cDNAs encoding a number of ovarian steroidogenic enzymes including the rainbow trout cholesterol side-chain cleavage cytochrome P-450, 3β-hydroxysteroid dehydrogenase (HSD), 17α-hydroxylase/17,20 lyase cytochrome P-450, aromatase cytochrome P-450 cDNAS as well as the pig 20β-HSD cDNA. Estradiol-17β stimulates the hepatic synthesis and secretion of a yolk precursor, vitellogenin. Vitellogenin is then transported to the ovary where it is selectively taken up into the oocyte by a receptor-mediated process involving specific cell-surface receptors. Estradiol-17β was also shown to induce the synthesis of egg membrane proteins in the liver. The maturation-inducing action of 17α,20β-DP and 20β-S is through the binding to the oocyte plasma membrane. This initial MIH-surface interaction is followed by the formation of the major mediator of MIH, maturation-promoting factor (MPF). We have purified MPF from mature oocytes of carp. Carp MPF consists of two components: the homolog of the cdc2⁺ gene product of fission yeast (p34^cdc2) and cyclin B. The cdc2 kinase protein is present in immature oocytes as well as in oocytes induced to mature by 17α,20β-DP treatment, while cyclin B proteins can be detected only in mature oocytes. Addition of bacterially expressed goldfish cyclin B to the extracts of immature goldfish oocytes induced MPF activation. These results suggest that the appearance of cyclin B protein is a crucial step for 17α,20β-DP-induced oocyte maturation in fish.     

Profiles of sex steroids, fecundity and spawning of a migratory characiform fish from the Paraguay–Paraná basin: a comparative study in a three-river system

This study investigated for the first time the reproductive biology of Prochilodus lineatus in a system of rivers in southeastern Brasil, relating it to the role of tributary rivers in the reproductive success of this important commercial fish in the Upper Paraná River basin, where a cascade of hydroelectric dams were deployed. Specimens were caught bimonthly in three river sites: (S1) Grande River, downstream from the Porto Colômbia dam; (S2) Pardo River; and (S3) Mogi Guaçu River. Sex steroid plasma levels, fecundity, follicular atresia, oocyte diameter and gonadosomatic index (GSI) were compared among sites. In S1, fish exhibited changes in the reproductive parameters: lower GSI, oocyte diameter and fecundity and higher follicular atresia index, when compared to S2 and S3. Frequency of maturing fish was higher in S3 and spawning was only registered in S3. In sites S2 and S3, plasma concentrations of testosterone and 17β-estradiol in females and testosterone in males showed wide variations following gonadal maturation. Fish from S1 showed few significant variations in sex steroid concentrations throughout the gonadal cycle. These results indicate that P. lineatus does not reproduce in Grande River (S1), but probably uses the Pardo River (S2) as a migratory route towards the Mogi Guaçu River (S3) where they complete gonadal maturation and spawning. Our findings contribute for understanding the reproductive biology of P. lineatus and to highlight the importance of tributaries in impounded rivers as a favourable environment for migration and spawning of fish.     

Indoor Spawning and Egg Development of the Red Claw Crayfish Cherax quadricarinatus

In 1991 and 1992, indoor hatchery experiments were conducted with red claw crayfish Cherax quadricarinatus a large tropical crayfish from northern Australia. Six fiberglass tanks were stocked with 40–190 g mature red claw crayfish at 10/m² and a 1:2 male to female ratio. Water temperature was maintained at 28 ± 1 C. Photoperiod was provided by natural daylight in 1991 and controlled lighting in 1992.
With natural lighting, monthly spawning rates (% of females spawning/month) ranged from an average of 15% during months with less than 12 h of light per day to over 35% in months with more than 12 h of light per day. Peak spawning occurred from May to July, too late to obtain juveniles for spring stocking. During the second year, photoperiod was increased from 10L:14D in December to 14L:10D in February and maintained at this level until July. Red claw spawning increased with increasing day length. Peak spawning rates of 50% per month and higher occurred from March to May. Spawning rates decreased dramatically after May, even though photoperiod and temperature remained constant. Eggs per spawn increased with an increase in female size. Fecundity average 7.3 juveniles per gram of body weight for females from 40 to 190 g, regardless of size. Egg color was useful for predicting stage of development.     

Influence of photoperiod and temperature manipulation on gonadal development and spawning in Caspian roach (Rutilus rutilus caspicus): Implications for artificial propagation

Caspian roach (Rutilus rutilus caspicus), a spring spawning teleost, were subjected to various photoperiod and temperature regimes to study the feasibility of shifting the timing of spawning for artificial propagation purposes. A total of 650 female reproductively mature R. rutilus caspicus were subjected to different photoperiod and temperature regimes including four light regimes (natural light (NL), 16 hr of light (L):8 hr of darkness (D), 9L:15D, 11L:13D), each affected by three temperature regimes (14, 20 and 24°C) for 70 days. Five fish per tank were randomly sampled on Feb. 10, Feb. 20, March 28, April 15 and April 30 (natural spawning time). Ovarian tissue sections were studied using light microscope and transmission electron microscope (TEM). The levels of 17‐β estradiol (E2) and 17α‐hydroxyprogesterone (OHP) were also measured in the serum samples. In late winter (March 28th), the gonadal maturation and spawning were accelerated in fish treated with the long day length (16L/8D) and warm temperature (20°C). While, the maturation of oocytes and spawning delayed in fish exposed to low temperature (14°C) and short day length (9L/15D and 11L/13D). Photoperiod seems to play a more important role in the ovarian development of the R. rutilus caspicus compared to temperature; since even among the fish treated with the lowest temperature (14°C), those exposed to a longer day length (16L/8D), matured and spawned earlier than the others. Considering that the earliest spawning occurred in R. rutilus caspicus treated with 16L/8D at 20°C and the latest spawning occurred in fish exposed to low temperature and short photoperiod, it can be concluded that temperature and photoperiod play an important role in accelerating oocyte maturation and spawning.     

Biosynthesis of estradiol-17β in the ovarian follicles of the red seabream Pagrus major during vitellogenesis

ABSTRACT: The red seabream Pagrus major is a useful experimental fish for studying the endocrine control of oogenesis in teleosts. This study investigated the steroidogenic pathway for estradiol-17β (E2) biosynthesis in the ovarian follicles of red seabream. Intact follicles were isolated during vitellogenesis and incubated in vitro with different radiolabeled steroid precursors. When 17-hydroxyprogesterone (17-P), dehydroepiandrosterone (DHEA), or androstenedione (AD) were used as precursors, both testosterone (T) and estrone (E1) were synthesized by follicles, leading to estradiol-17β (E2) production. Serum steroid levels measured by enzyme-linked immunosorbent assay showed that T, E1, and E2 were present in the circulation at levels ranging from 1 ng/mL to 2 ng/mL throughout the day during the spawning season. In vitro conversion of E1 into E2, however, was 15.8-fold greater than T conversion into E2, suggesting that E2 is synthesized mainly via E1 rather than T. The results showed that E2 was synthesized from pregnenolone via 17-hydroxypregnenolone, DHEA, AD, and E1. Thus, the study demonstrated the complete steroidogenic E2 synthesis pathway in the ovarian follicles of red seabream, and revealed that E1 is the major precursor of E2.     

Conditioning,maturation and year‐round natural spawning of orange‐spotted grouper,Epinephelus coioides (Hamilton, 1822), in recirculating aquaculture system

The present experiment was aimed at studying the conditioning, maturation and natural spawning of orange‐spotted grouper, Epinephelus coioides, in a recirculatory aquaculture system (RAS). Thirty fish (n = 30; 3.35 ± 0.05 kg) were stocked in a circular tank of 125 m³ capacity fitted with an RAS for conditioning and broodstock development. After 15 days, 15 fish were implanted with 17 α methyl testosterone and letrozole at the rate of 5 mg and 0.2 mg/kg body weight, respectively, for conversion from female to male. The gonadal development started after 1 month, and by 90th day, 63.53 ± 3.78% and 2.07 ± 0.84% of the oocytes attained a size of 400–500 μm and 500–600 μm respectively. Natural spawning commenced in the RAS from 4th month onward after stocking and spawning continued round the year. The spawning pair showed courtship behaviour with a typical vertical burst of swimming just before release of gametes. The total number of eggs spawned during 1 year was 47.23 million with spawning frequency varying form 5 to 13 times per month. The association of spawning events with new moon day (lunar cycle) weakened as time progressed. The mean monthly fertilization and hatching rates varied from 77.80 ± 3.34% to 83.70 ± 1.76% and 82.80 ± 4.21% to 88.33 ± 1.39% respectively. The study proved that RAS is an efficient system that provides a stable, controllable and conducive environment for year‐round natural breeding of orange‐spotted grouper.     

Sex reversal in the genus Oreochromis: 1. Immersion of eggs and embryos in oestrogen solutions is ineffective

Failure to achieve feminization of tilapia under conditions where masculinization is successful was attributed to earlier determination of testis development than of ovary development. Success in obtaining sex inversion by immersion of eggs and fry of salmonids in hormone solutions suggested the possibility of obtaining sex inversion by treatments applied at early life stages. Thence, tilapia embryos were exposed to oestrogens as early as possible after fertilization, by incubating the eggs in oestrogen solutions. Further treatments consisted of rearing the fry in oestrogen solutions for various durations. Eggs of Oreochromis mossambicus (L.) and of O. mossambicus×O. urolepis hornorum Trewavas were collected from the mouths of females immediately after spawning and incubated in Erlenmeyer flasks set in a water bath shaker, at 27°C. The following oestrogenic hormones (dissolved in ethanol) were added to the flasks: 17β-oestradiol, progesterone, flutamide, and flutamide + progesterone. Initial treatments were given at concentrations of 1-50αg/l, for different durations ranging from 7 to 30 days after spawning. Due to high mortalities, higher doses (i.e. 100-5000αg/1) were tested for shorter periods (from 2 hours to 12 days). No sex inversion to female or to intersex gonads was achieved in any of the immersion treatments.     

17β-雌二醇对双齿围沙蚕幼虫SOD、CAT和GST活性的影响

将双齿围沙蚕（Perinereis aibuhitensis）幼虫暴露于不同质量浓度的17β-雌二醇（0.1μg·L^-1、1μg·L^-1、10μg·L^-1、100μg·L^-1和1 000μg·L^-1）中,于第2、第4、第6和第8天取样,分别测定抗氧化酶[超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和谷胱甘肽硫转移酶（GST）]活性。结果表明,第2天时10μg·L^-1浓度组显著诱导双齿围沙蚕CAT、SOD和GST活性,1 000μg·L^-1浓度组显著抑制其SOD和GST活性。第4天时各浓度组SOD和GST活性均被不同程度诱导。第6天时低浓度组（0.1μg·L^-1和1μg·L^-1）显著抑制双齿围沙蚕CAT活性,高浓度组（100μg·L^-1和1 000μg·L^-1）则抑制其SOD活性。第8天时,除1 000μg·L^-1浓度组外,其余各组CAT活性均被不同程度诱导（P〉0.05）,而SOD活性则受到抑制,10μg·L^-1浓度组显著抑制其GST活性。试验结果显示,外源性17β-雌二醇对双齿围沙蚕幼虫产生氧化胁迫,沙蚕幼虫SOD、CAT、GST对17β-雌二醇的响应与其暴露浓度及时间有关。     

Larviculture of the Greasy Grouper Epinephelus tauvina F. and the Brown-Marbled Grouper E. fuscoguttatus F. in Singapore

This paper reports the larviculture of two grouper species, the greasy grouper Epinephelus tauvina and the brown-marbled grouper E. fuscoguttatus , and examines the technical feasibility of breeding the fish. Fertilized eggs for larviculture were obtained by induced spawning through multiple hormonal injections of the female brooders, followed by artificial fertilization for the greasy grouper and by natural spawning in netcages for the brown-marbled grouper.
For both species, larviculture was divided into three operational stages, i.e., stage 1 (day 0–12), stage 2 (from day 12 to day 24 for the greasy grouper and to day 30 for the brown-marbled grouper) and stage 3 (day 24 or 30 to metamorphosis). Larvae were transferred to clean tanks after each stage. Total mortality around day 5–8 was common for the greasy grouper. Several modifications, including the feeding with super-small strain rotifers, intensive feeding of rotifers with Nannochloropsis and the use of an oil-skimmer, have been made to improve the situation. Two other major problems in larviculture were the high mortality observed after day 25 in the greasy grouper and the high cannibalism from day 35 onwards for the greasy grouper and from day 30 onwards for the brown-marbled grouper. The shock syndrome exhibited by both species during the periods made culling of the fish impossible.
Based on the spawning and larval characteristics, brown-marbled grouper is considered a better potential species for large scale fry production than the greasy grouper.     

Seasonal changes in plasma gonadal steroid concentrations and gonadal morphology of male and female tench (Tinca tinca, L.)

In order to gain a better understanding of the reproductive cycles of male and female tench (Tinca tinca), gonadosomatic index, gonad histology and plasma concentrations of estradiol‐17β (E₂), testosterone, an drostenedione, 11‐ketotestosterone (11‐KT), 17,20β, 21‐trihydroxy‐4‐pregnen‐3‐one (17,20β,21‐P), 17,20β‐dihydroxy‐4‐pregnen‐3‐one (17,20β‐P) and 17,20α‐dihydroxy‐4‐pregnen‐3‐one (17,20α‐P) were measured at the four seasons of the year, plus a further sampling coincident with the peak of spawning in early July. As expected, in both males and females, the plasma concentrations of androgens (excluding 11‐KT in females – undetectable) and C₂₁ steroids were significantly more elevated in the spring and summer (when most gonadal development took place) than in the autumn and winter. The only unexpected finding was that 17,20β‐P and 17,20β,21‐P, the steroids that are normally associated with oocyte final maturation in females and spermiation in males, were found in substantial amounts in both pre‐vitellogenic, pre‐spermatogenic and post‐spawning fish. This suggests that these steroids may have other as yet unidentified roles in this species.     

Characterization and comparison of variations in reproductive performance of Chitralada strain Nile tilapia, Oreochromis niloticus (L.)

In an attempt to better understand and characterize the variability in the female reproduction of individual Thai‐Chitralada strains of Nile tilapia, Oreochromis niloticus (L.), tagged mature females (n=68) from a single population were monitored in a hapa‐in‐pond system over a 12‐month experimental period. Spawn female⁻¹, eggs female⁻¹ day⁻¹, eggs kg female⁻¹ day⁻¹, day spawn⁻¹ and weight (g) at first spawning of individual females were determined from the regular weekly sampling of weight (g) and eggs per spawn. For analysis, the females were grouped into two, high spawning (HSF) and low spawning frequency (LSF) classes based on their spawning frequency (SF) record. Moreover, nested under these two classes were two groups each based on growth rate, i.e., high frequency – large size (HL), high frequency – small size (HS), low frequency – large size (LL) and low frequency – small size (LS). There was no difference in eggs spawn⁻¹ among all females. The HSF group produced 68% and 361% more eggs female⁻¹ day⁻¹ than the population mean and LSF group respectively. Eggs female⁻¹ day⁻¹ and spawn female⁻¹ remained high in the HSF group and low in the LSF group throughout the 12‐month experimental period. This suggests that individual female spawning activity is consistent within a population in a common environment. The inter‐spawn interval increased with age in all four groups, and day spawn⁻¹ was shorter by 130% in HSF females compared with day spawn⁻¹ in LSF. The HSF group also spawned more successively (≥3), while the LSF group of females had fewer successive spawns (≤2). Body weight (g) had no influence on the number of eggs produced. The study indicates that separating frequently spawning females could be used as an important strategy to improve commercial seed production of Nile tilapia.