Reproductive ability of triploid ornamental (koi) carp (Cyprinus carpio L.) × goldfish (Carassius auratus L.) hybrids and characteristics of their offspring

The purpose of this study was to investigate reproductive ability of backcross triploid koi (Cyprinus carpio L.) × goldfish (Carassius auratus L.) hybrids. These triploids have been obtained by crossing of F₁ hybrid females producing diploid eggs with males of parental species. Triploid hybrid females, when crossed with goldfish or koi males, produced mostly aneuploid fish with ploidy range from approximately 2.2n–3.2n with a mean value 2.5n; some fish in crosses of triploid females with koi males were tetraploid (4.0n). Since analysed fish had in their genomes one haploid set from parental males, the data indicate that triploid hybrid females mostly produced aneuploid eggs with ploidy range from approximately 1.2n–2.2n and a mean ploidy around 1.5n while some eggs were triploid. Triploid hybrid males were completely sterile and have not released any sperm after hormonal injection. Despite their low viability, some aneuploid fish obtained from triploid hybrid females were raised in indoor recirculating systems until the age of 2 years and their reproductive ability has been evaluated. One aneuploid female with ploidy 2.1n produced larvae with ploidy range from 2.9n to 3.4n with a mean ploidy of 3.1n when crossed with a koi male; about 60% of obtained larvae had ploidy from 3.0n to 3.2n. These data indicate that this female produced mostly eggs with unreduced ploidy level.     

Reproductive ability of second generation ornamental (koi) carp (Cyprinus carpio L.) × goldfish (Carassius auratus L.) hybrids and characteristics of their offspring

The purpose of this study was to investigate the reproductive ability of second generation (F₂) koi (Cyprinus carpio L.) × goldfish (Carassius auratus L.) hybrids. Only diploid F₂ males and females were fertile and used in crosses. A significant increase was recorded in male fertility in F₂ versus F₁. In contrast with an earlier study in which only one fertile F₁ male was found, about 20% of F₂ males produced sperm. The observed reproductive ability of F₂ hybrids was similar to that demonstrated by the only fertile F₁ male and F₁ females. F₂ males produced diploid spermatozoa and generated triploids when crossed with koi females. All triploid fish in these progenies were males indicating that F₂ males had a sex chromosome constitution of XY. F₂ females produced diploid eggs and generated mostly triploids when crossed with koi males. In progenies obtained by crosses of F₂ males with F₁ and F₂ females, most of the surviving juveniles (63%–100%) were diploid; a minority of juveniles were aneuploid (ploidy ranged from 2.1n to 3.6n). Diploid fish in these progenies were presumably the result of spontaneous androgenesis and gynogenesis, by the same mechanisms observed earlier in progenies obtained by crossing the F₁ male with F₁ females.     

Ovaprim treatment promotes oocyte development and milt fertilization rate in diploid and triploid African catfish (<Emphasis Type="Italic">Clarias gariepinus)</Emphasis>

Triploid fish are increasingly used in aquaculture because they are generally unable to reproduce successfully. Energy is channeled into somatic growth rather than gonadal development, and in the event of escape, the animals are unlikely to breed successfully among themselves or with wild conspecifics. This study tested the ability of recently matured triploid African catfish (Clarias gariepinus) to produce and fertilize eggs with and without ovaprim treatment. Triploid females did not show the increase in ovary size observed in diploid members of the same cohort between 8 and 9 months of age, or the coincident decrease in visceral fat deposits, and this was unaffected by up to 5 weekly i.m. injections of 0.5 ml kg⁻¹ Ovaprim. However, we observed advanced vitellogenin (Vtg) sequestration in oocytes of triploid females, albeit to a lesser degree and with lesser cortical alveoli, compared to oocytes from diploid cohort members. Histological sections revealed a positive trend of oocyte development up to the third weekly ovaprim injection followed by a negative gonadal development in weeks four and five. Milt from triploid males injected 9–12 h earlier with 0.25 ml kg⁻¹ ovaprim i.m. fertilized more diploid eggs than milt from untreated triploid males (30 vs. 20%), but none of the developing embryos of triploid paternity survived to hatch. In contrast, milt of diploid males fertilized 49% of eggs, and 20% of the developing embryos hatched successfully. These rates were improved in ovaprim-injected diploid males to 70% fertilization and 33% hatch. This study demonstrates potential of overcoming non-viability of eggs from triploid female African catfish, and enhancing the ability of triploid milt to fertilize eggs.     

Induction of meiotic gynogenesis in the stinging catfish Heteropneustes fossilis (Bloch) and evidence for female homogamety

This study reports the results on induced meiotic diploid gynogenesis and female homogametic nature in the Indian catfish, Heteropneustes fossilis. The eggs of H. fossilis were inseminated with conspecific sperm. The sperm suspension was diluted to 1 × 10⁷ sperm mL⁻¹ in Hanks balanced salt solution. Sperm were irradiated under UV light, with the exposure time ranging from 15 to 360 s (7500 ergs mm⁻² for 60 s). The genetic inactivation of paternal chromosomes was confirmed by chromosome counting from the larval cells and the larvae also had a characteristic haploid syndrome. A typical ‘Hertwig effect’ in the yield of hatched larvae was observed with doses of UV exposure >75 s (9375 ergs mm²). Larvae resulting from sperm UV irradiated above 120 s (15 000 ergs mm²) were 100% haploids. Application of heat shock to the activated eggs was effective in suppressing the release of the second polar body (meiotic gynogenesis) and resulted in diploid gynogenetic larvae morphologically identical to those of the control. The best yield of diploid gynogens (49.3% with respect to the control) was found to be at 6 min after egg activation and the heat shock at 41 °C for a 1-min duration, at an ambient water temperature of 27 °C. A total of 113 diploid gynogenetic fry from seven different female fish were reared and subjected to sexing. All gynogenetic fish were female in contrast to the control, which had a mean sex ratio of 56.7% females (which was not significantly different from 50% female). From these results, the sex determination mechanism in H. fossilis was presumed to be female homogamety.     

The effect of individual male potency on fertilization ability of fresh and cryopreserved milt of rainbow trout, Oncorhynchus mykiss (Walbaum)

The goal of the present study was to examine individual male potency in rainbow trout, Oncorhynchus mykiss (Walbaum), expressed as the fertilization ability of fresh and cryopreserved sperm. One female and four males bearing genetic markers enabling determination of the progeny paternity were chosen as gamete donors. Samples of eggs were inseminated with sperm from separate individuals or with pooled sperm. Genetic examination of the progeny obtained after fertilization of eggs with pooled milt showed differences in male potency. The proportions of offspring sired by four individual males after fertilization of eggs with the fresh milt were similar to those obtained after fertilization with cryopreserved milt (correlation r = 0.95; n = 4; P < 0.05). These proportions did not correlate with the proportions of progeny resulting from fertilization of eggs when sperm was not pooled.     

The breeding potential and biochemical composition of triploid Manila clams, Tapes philippinarum Adams and Reeve

In 1989 and 1990. triploid Manila clam, Tapes philippinarum Adams and Reeve, seed were reared to 15-20 mm at the Fisheries Laboratory, Conwy, and planted out in the Menai Strait, North Wales. In each of the summers of 1992 and 1993, three of these batches, at 2, 3 or 4 years old, were returned to the laboratory to assess ploidy, size, spawning potential and biochemical composition. Percentage triploidy at this time was similar to that in the seed. After 6 and 8 weeks of warmwater conditioning. Only 45 out of l21 triploids (37%) were induced to spawn by thermal shock, with only one spawning as a male. By comparison, 75% of diploid clams spawned with a 1:1 ratio of males to females. Mean fecundity of triploids was significantly lower than that for diploids, at 0.497 compared with 1.54 million eggs per female. Compared with eggs from diploid females, eggs from triploids were larger and significantly fewer of them developed into D-larvae when fertilized by sperm from diploid males. Triploid clams were heavier and had a higher condition index and carbohydrate content than diploids of the same age, but lipid levels were similar. Potential advantages of producing and cultivating 100% triploid batches of Manila clam seed are discussed.     

Induction of triploidy in mud loach (Misgurnus mizolepis) and its effect on gonad development and growth

Triploidy was induced in mud loach (Misgurnus mizolepis) by cold shocking fertilized eggs 5 min post-fertilization at 2°C for 15 to 60 min. Best results were obtained when eggs were shocked for 60 min; 98% of fish examined in that treatment were triploids. Triploidy was confirmed by erythrocyte measurements and chromosome counts. Diploids had 48 chromosomes, while triploids had 72. Histological analysis of 9-month-old triploid ovaries showed an appreciable number of oocytes at the chromatin nucleolus stage with considerable interstitial tissue. However, diploids had well developed oocytes. Diploid testes from diploid males exhibited normal spermatids and spermatozoa, while a few were seen in triploid males. Growth rate was evaluated over a 9-month growth trial. Although male and female triploids were slightly heavier than their diploid counterparts from the third to the ninth month, their growth rates were not significantly different compared to their diploid controls.     

Genetic contribution of parents in sex determination of honmoroko Gnathopogon caerulescens

The honmoroko has been inferred to have an XX/XY sex determination system, but the parental genome can also affect the sex ratio of the offspring. The extent of parental effects on sex determination was examined by checking the sex ratios of F1 and F2 gynogenetic diploids and control diploids. Eleven gynogenetic broods from different females consisted of all or nearly all females, but eight broods showed a variable proportion of males (<50 %). One second-generation brood of gynogenetic diploids consisted wholly of females, but others produced some males. In crosses with a control diploid female, four males from a high-percentage male brood of gynogenetic diploids produced offspring with a balanced sex ratio. Sib-mating between a gynogenetic female and three gynogenetic males from the brood produced predominantly male progeny. These results suggest that there are at least four possible genotypes: genotypic female (XX), phenotypic female carrying a silent Y chromosome, genotypic male (XY), and genotypic supermale (YY). These inferences suggest that this fish has an XY system but a relatively high proportion of females possess a mutated, silent Y chromosome which does not lead to testis formation.     

Gynogenesis induction and sex determination in the Eurasian perch,Perca fluviatilis

In the present study, we used meiotic gynogenesis, widely used in studies on sex determination, to confirm female homogamety in Eurasian perch, Perca fluviatilis. Sperm irradiated with UV for 400 s was used to artificially fertilized eggs. The diploid of the resulting embryos was restored by a heat shock (30 °C) applied to the eggs 5 min postfertilization, for 25 min. Fertilization (ranging between 45% and 75%) and survival rates at hatching (ranging between 3.4% and 46.6%) were not significantly different (P>0.05) between the diploid control and gynogenetics. The diploid controls and two batches of gynogenetics contained 100% diploid larvae, whereas two other batches of gynogenetics contained 6.7% and 10.0% triploid larvae. The sex ratios of the diploid controls were not significantly different from 1:1, whereas all gynogenetic families were 100% female. These results confirm female homogamety in Eurasian perch, demonstrated by the use of hormonally mascilinized breeders in a previous study.     

Effects of temperature and salinity on the reproductive success of Arctic charr, Salvelinus alpinus (L.): egg composition, milt characteristics and fry survival

The effects of temperature and salinity on the reproductive success of Arctic charr, Salvelinus alpinus (L.), were examined by holding broodstock under the following conditions from mid‐May until the end of September: fresh water at ambient temperature (NFW; 8–16 °C); salt water (25–30‰) at ambient temperature (NSW; 4–10 °C); fresh water cooled to saltwater temperature (CFW; 4–10 °C); or salt water heated to freshwater temperature (HSW; 8–16 °C). The relative fecundity of females was similar among groups (P > 0.05; 2685 ± 706 eggs), but females reared in NSW produced significantly larger eggs than those raised in NFW. The highest spermatozoa concentrations were found in milt from males reared in SW and the highest milt glucose concentration was from males kept under coldwater conditions (CFW, NSW). Eggs from NSW and HSW females contained more proteins than eggs produced by NFW females. Eggs from NSW females also contained 40% more lipids than was observed in the other groups, and total energy content was 27% higher in eggs from NSW females than in eggs from NFW females. When FW was cooled (CFW), females produced eggs with protein contents similar to those in NSW, but the lipid contents remained 30% lower. Finally, the best survival at the eyed stage and at hatch was observed in families produced by NSW broodstock. Intermediate values were observed in families from NFW or CFW while the highest mortality occurred in families from the HSW group. All these results suggest that, under the experimental conditions used in the present study, coastal seawater conditions offered the most favourable summer rearing conditions with respect to the reproductive success of Arctic charr.     

Genetic certification of presumed hybrids of blue × red abalone (Haliotis fulgens Philippi and H. rufescens Swainson)

Presumed hybrids between Haliotis rufescens and H. fulgens abalones, produced in a commercial abalone laboratory, were evaluated for progeny viability, gonad characteristics and for their ‘true’ status as hybrids by means of allozyme and microsatellite analyses. Four out of 10 evaluated allozyme loci proved to be useful for the genetic certification of hybrids, Gpi, Mdh‐1, Mdh‐2 and Sod, as they had exclusive alleles for each of the abalone reference species from which the presumed hybrids were produced. Three microsatellite loci were also evaluated for their usefulness in abalone hybrid certification. Two were not useful for hybrid certification as the reference abalone species shared some of the same alleles. The third loci (Hka 56) also had shared alleles between the two species, but at very different frequencies. Therefore, this permitted a presumptive certification of hybrids, which, however, was not error‐proof as two allozyme certified hybrids were wrongly classified as H. rufescens rather than as hybrids. Progeny viability of the only female certified to be a hybrid was significantly lower compared with one viability of the other females spawned, which were certified as H. rufescens. However, the number of eggs spawned by the only certified hybrid female was as large as those spawned by females certified as H. rufescens. Gonads of abalones sampled for the macroscopic appearance of sterility indicated that in the absence of any gonad development, no distinctive difference allows for hybrid identification, as there were undifferentiated individuals that were both hybrid and non‐hybrid. On the other hand, two certified hybrid males were characterized by spermatocytes being the most abundant gamete type, although one had some spermatozoa present, indicating only partial sterility. Among the two female hybrids found, vitellogenesis was observed to proceed up to vitellogenic oocytes.     

Study of sex determination system in ship sturgeon, Acipenser nudiventris using meiotic gynogenesis

The sex ratios and sex determination mechanism of gynogenetic diploids of ship sturgeon, Acipenser nudiventris, have been investigated to verify the possibilities of sex control by chromosome manipulation in this species. Meiotic gynogenesis was induced in a female ship sturgeon using cold shock after egg activation with UV-irradiated sperm of a male Siberian sturgeon. Microsatellite DNA analysis was applied for verification of uniparental inheritance in the gynogenetic diploid group of fish. All the analyzed gynogenetic diploids possessed only maternal genotype in the examined experimental group of fish. In this study, a minimum of two distinctly selected diagnostic loci in the offspring was used to confirm exclusively maternal contribution. Also, these fish were analyzed for sex diagnostic. Histological analysis of gonads from gynogenetic diploids, obtained from one family, showed 73.3 % of females and 27.7 % of males. The observed sex ratio has suggested that the ship sturgeon have a female heterogametic sex determination system. Gynogenesis in this species with female heterogametic sex determination system will have important role in breeding program and reclamation of its natural population to produce both female and male progeny, while this species has been introduced in the red list of IUCN (International Union for Conservation of Nature and Natural Resources).     

Early out-of-season induced spawning of channel catfish Ictalurus punctatus (Rafinesque) conditioned in heated earthen ponds

This study documents early out-of-season induced spawning of channel catfish Ictalurus punctatus. During the early spring (February–April) of 1999, 2000 and 2001, ponds containing (1) male and female channel catfish (mixed-sex ponds) or (2) male channel and blue catfish I. furcatus only, or female channel catfish only (single-sex ponds) were heated to 24–30°C to encourage gonadal maturation and spawning. Unheated ponds were stocked with males and females and were monitored during the duration of heating. When natural spawning occurred in the heated ponds, the fish were captured by seining and unspawned females were injected with 100 μg kg⁻¹ of synthetic leutenizing hormone-releasing hormone. Injected females were either paired with males or held in communal all-female groups, and monitored for ovulation. Eggs were collected and fertilized with sperm of channel catfish or blue catfish. Females paired with males were induced to spawn 44 days (mixed-sex ponds) and 50 days (single-sex ponds) before natural spawning occurred in unheated ponds. Spawning latency (the time between injection and ovulation) and the percentage of neurulated embryos from eggs fertilized using channel catfish sperm was not different between spawning before the natural season (P=0.68) and during the natural season in fish from mixed-sex ponds (P=0.57). Females held in all-female groups produced eggs 34 days before the onset of spawning in unheated ponds. Spawning latency was not different between spawns before and during the natural season (P=0.16), and the percentages of neurulated embryos from eggs fertilized with channel catfish sperm (P=0.76) or blue catfish sperm (P=0.77) before or during the natural season were not different. This study demonstrates the feasibility of conditioning of channel catfish females for early out-of-season induced spawning in the laboratory.     

Hormonal profile of growing male and female diploids and triploids of the blue tilapia,Oreochromis aureus,reared in intensive culture

Triploidy as a result of thermal shock exposure of fertilized eggs decreases the growth rate ofOreochromis aureus as compared to their diploid controls, but this is due to the higher female ratio present in triploids (86%) and the lower growth rate of females. When females and males are considered separately, the growth rate is not significantly different in diploids and triploids. Since triploidy results in a malfunctioning steroidogenesis in females (mainly testosterone (T) and 17β-estradiol (E₂)), but does not affect the growth rate, it is concluded that female gonadal steroids do not influence growth unless in pharmacological concentrations. These low levels of gonadal steroids are generally accompanied by higher levels of gonadotropin (GtH), but the difference is not always significant. Despite their lower growth rate diploid females have higher plasma concentrations of growth hormone (GH) during several months compared to the triploid females and diploid males. 3,5,3′-triiodo-L-thyronine (T₃) levels, however, are comparable between diploid and triploid females (except for 1 month), but higher in diploid males in 4 of the 5 months studied. 11-ketotestosterone (11kT) is always higher in males. These results indicate that the higher growth rate of males may be related to the high circulating levels of T₃ and 11kT.     

All-male Tilapia Hybrids of Two Strains of Oreochromis niloticus

Hybridization between Nile tilapia Oreochromis niloticus (Local and Stirling University strains) and blue tilapia O. aureus was found to be a tool to produce monosex populations. In order to select a purebred that produces all- or nearly all-male hybrids with high productivity, O. niloticus females and their diploid gynogenetics (meiogynes and mitogynes) were hybridized with O. aureus males. The sex ratio of progenies was evaluated from inter- and intraspecific crosses of two strains of Nile tilapia. Single-pair matings and group spawns under hatchery conditions showed no deviation ( P > 0.01) from the expected sex ratio of intraspecific crosses among two strains of Nile tilapia. However, a higher proportion of male progenies in blue tilapia was observed in group spawns in pond ( P < 0.004) and hatchery conditions ( P < 0.01). Only the Stirling strain and mitogynes produced all-male progenies under laboratory conditions. There were significant differences ( P < 0.05) in growth among hybrid progeny groups, when gynogens and their regular O. niloticus (Local strain) females were crossed with O. aureus males. Six-month hybrid offspring from mitogyne female parents grew better than those from regular and meiogyne female broods.     

Genetic characterization of the progeny of a pair of the tetraploid silver crucian carp Carassius auratus langsdorfii

Silver crucian carp Carassius auratus langsdorfii comprises a diploid-polyploid complex in wild Japanese populations. Bisexually reproducing diploids are sympatrically distributed with gynogenetically developing triploids and tetraploids. Triploid and tetraploid males are very rare among Japanese silver crucian carp due to their gynogenetic reproduction. We examined the genetic characteristics of progeny that arose in a tank by natural spawning of a tetraploid silver crucian carp pair. The ploidy status of 120 samples randomly collected from these progeny was determined to be tetraploid by DNA content flow cytometry. DNA fingerprints from a random amplified polymorphic DNA assay indicated that almost all the progeny examined had genotypes identical to the maternal tetraploid female with no paternally derived fragments. Selected specimens’ cytogenetic analyses revealed that the progeny examined had tetraploid chromosome numbers, categorized into 40 metacentric, 80 submetacentric, and 80 subtelocentric or telocentric chromosomes, which were arranged into quartets and six supernumerary microchromosomes. Fluorescence in situ hybridization signals were detected in four homologous chromosomes in all analyzed metaphases prepared from diploid goldfish specimens. Contrary, tetraploid silver crucian carp gave eight rDNA signals. These results suggest that gynogenetic development in eggs spawned by tetraploid females should be triggered by tetraploid males’ homospecific sperm.     

Reproductive performance and mature gonad morphology of triploid and diploid Black Tiger shrimp (Penaeus monodon) siblings

Sibling harvest age Black Tiger shrimp triploids and diploids of both sexes were reared to reproductive maturity, crossed with wild caught females and males, conditioned for spawning and a comprehensive reproductive performance trial was undertaken. Ovarian development, spawning frequency, fecundity, hatch rate, gonad morphology, male reproductive tracts and thelycum impregnation rates of the wild female × triploid male cross were assessed. After ablation, ovarian development and cycling between wild G₀ diploid and G₁ diploids was not significantly different, whereas G₁ triploids failed to show any signs of ovarian development and cycling, thus resulting in no G₁ triploid female spawnings. There were 10 G₀ diploid female × G₀ diploid male first‐spawnings and 9 G₀ diploid female × G₁ diploid male first‐spawnings, all of which produced viable nauplii. In comparison, there were 7 G₀ diploid female × G₁ triploid male first‐spawnings, none of which produced viable nauplii. The 26 wild G₀ diploid female spawnings had more eggs than the 1 G₁ diploid female spawning. Gonad morphology and male reproductive tract assessments showed impaired reproductive development in triploid gonadal tissues of both sexes (compared with sibling diploids and wild shrimp) to a point where complete maturation had not occurred. The thelycum of 16 wild G₀ diploid females crossed with G₁ triploid males had no visible spermatophore present, suggesting that G₁ triploid males are incapable of developing viable spermatophores and mating with females. This study demonstrates that the triploid females and males are incapable of producing viable gametes and are thus reproductively sterile.     

Triploid hard clams Mercenaria mercenaria produced by inhibiting polar body I or polar body II

Two types of triploid hard clams Mercenaria mercenaria were produced by inhibiting polar body I (PB1) or polar body II (PB2) with cytochalasin B. Treatments were applied at 22–23°C, with PB1 inhibition starting at 4–7 min postfertilization and ending when PB2 was first observed in control groups, and with PB2 inhibition starting at 17–23 min postfertilization and ending when 80% of control eggs released PB2. Triploid induction success was evaluated by chromosome counting in 2–4 cell embryos and by flow cytometry at larval and juvenile stages. PB2 inhibition produced more triploids (82%–100%) than PB1 inhibition (71%–83%), although the difference was not significant (p ≥ .088). Triploid percentages in PB1‐ or PB2‐inhibited groups showed a small but insignificant decline during the first 6 months. At month 3, PB1 and PB2 triploids were not different from their within‐group diploids, but significantly larger than control diploids; PB1 triploids were significantly larger than PB2 triploids (p ≤ .003). At month 6, PB1 triploids were not different from either within‐group or control‐group diploids, while PB2 triploids were significant larger than both within‐group and control diploid; PB1 triploids were smaller than PB2 triploids. At month 16, PB1 and PB2 triploids in one remaining replicate were not different from their within‐group diploids. Overall, this study shows that triploids can be efficiently produced by PB1 or PB2 inhibition, and their growth performance relative to diploids is variable depending on age and replicates or parental genotype.     

Allelic segregation of sex‐linked microsatellite markers in Nile tilapia (Oreochromis niloticus) and validation of inheritance in YY population

Stocking of all‐male fingerling produced by direct administration of male hormone 17‐α‐methyltestosterone is the most preferred method for present‐day aquaculture of the Nile tilapia Oreochromis niloticus. However, due to the growing concern of negative impact of steroid hormone in food fish, production of ‘genetically male’ tilapia, which depends on the concrete and thorough understanding of sex determination, has long been a scientific curiosity. The objective of the present study was to identify reliable sex‐linked markers and to evaluate the applicability of those markers in terms of monosex production approach. ‘XY’ neofemales were produced by using synthetic oestrogen and identified through selective breeding and progeny testing. Three females with progeny not deviating from 3:1 sex ratio (male:female) were designated as ‘XY’ neofemales and were used subsequently to produce putative YY progeny. Among the fifteen microsatellite markers tested, marker ARO172 was most informative in differentiating male and female genotypes. Twenty‐seven F₂ fish from three families were identified as putative YY males based on marker genotyping, and four of them were crossed to produce F₃ to validate marker association by progeny testing. The YY males produced 86%–100% male progeny indicating ARO172 a unique sex‐linked marker applicable in marker‐assisted selection.     

Sperm from pheromone primed brown trout (Salmo trutta L.) produce more larvae

Male goldfish (Carassius auratus) exposed to female hormonal pheromones express increased milt volumes and their sperm fertilize more eggs than sperm from unprimed males. Ovulated salmonid females also release odours that increase volumes of strippable milt in males. It is, however, not known if the priming pheromones affect the ability of sperm to fertilize eggs in salmonids. In this study, we compare the proportion of larvae produced from in vitro fertilization tests between primed brown trout (Salmo trutta) males exposed to a mix of female urine and ovarian fluids, and control males exposed only to 0.9 % sodium chloride. We also investigate priming effects on milt yield and sperm motility. Fertilization tests with sperm from single males, as well as sperm from two males (i.e., sperm competition), were performed. Primed males generated more larvae in both the single male and competition fertilization tests. No differences between treatments in milt yield and sperm motility could be established.