Hormonal and pheromonal control of spawning behavior in the goldfish

Species that employ sexual reproduction must synchronize gamete maturity with behavior within and between genders. Teleost fishes solve this challenge by using reproductive hormones both as endogenous signals to synchronize sexual behavior with gamete maturation, and as exogenous signals (pheromones) to synchronize spawning interactions between fish. This dual role of hormonal products is best understood in the goldfish, an external fertilizer with a promiscuous mating system. Female gonadal growth and vitellogenesis is stimulated by 17β-estradiol (E2) which also evokes release of a recrudescent pheromone. At the completion of vitellogenesis, ovarian E2 production drops and plasma testosterone increases, sensitizing the female gonadotropin II (luteinizing hormone; LH) system to environmental cues (temperature, spawning substrate, pheromones). These cues eventually trigger a LH surge that alters steroidogenesic pathways to favor the production of progestins including 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P). Plasma 17,20β-P stimulates oocyte maturation but is also released to the water along with sulfated 17,20β-P and androstenedione to serve as a preovulatory pheromone. This pheromone stimulates male behavior, LH release, and sperm production. At the time of ovulation, females become sexually active in response to prostaglandin F2α (PGF2α) synthesized in the oviduct. PGF2α and its metabolites are released as a postovulatory pheromone that induces male spawning behavior which further increases male LH and sperm production. Androgenic hormones are required for male behavior and LH release. Although goldfish are gonochorists, hormone treatments can induce heterotypical functions in adults. Similar findings in other fish demonstrate that a sexually bipotential brain is not restricted to hermaphroditic fishes. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Gonadal recrudescence and induced spawning in Barbus altianalis

Gonadal recrudescence in Barbus altianalis from River Nile, Jinja (33°05E; 0°45N) Uganda was studied prior to induced spawning. Oocytes in B. altianalis were found to develop in cohorts suggesting batch spawning throughout the year with pronounced spawning activities coinciding with rainfall peaks of April and September. There was a strong positive correlation between rainfall and gonadosomatic index (r_S = 0.75, P = 0.008). B. altianalis had low fecundity (6.0 ± 2.0 per gram) and large‐sized eggs (2.97 ± 0.1 mm). Induced spawning was successful with exogenous hormones and water current. Results from this study suggested that total striping may not be appropriate, but larger eggs would confer high survival of offsprings under optimal conditions. The success registered in induced spawning of B. altianalis provides a breakthrough in seed production for multiplication and culture of this economically important fish species.     

Evaluation of cGnRH IIa for induction spawning of two ornamental Epalzeorhynchos species

The administration of exogenous hormones for induction spawning of captive fish is a common practice throughout the finfish aquaculture industry. Induced spawning protocols for ornamental cyprinids commonly rely on sGnRH IIIa, commercially available as Ovaprim. In this study, the efficacy of an alternative spawning aid, cGnRH IIa, was evaluated relative to Ovaprim, in two commonly cultured ornamental Cyprinids: the Redtail Sharkminnow Epalzeorhynchos bicolor and the Rainbow Shark E. frenatum. Broodstock of each species were injected with either a positive control (0.5 µl/g Ovaprim), a negative control (propylene glycol) or one of three doses of cGnRH IIa (50, 100, or 200 µg/kg). Following spawning aid injection, ovulation success, fecundity, fertilization success, embryo diameter, hatch success and larval notochord length were evaluated. Ovulation success (82 ± 8%) was statistically similar to the positive control group for all experimental doses of cGnRH IIa in E. bicolor, while 50 and 100 µg/kg cGnRH IIa doses resulted in significantly higher ovulation success (100.0% and 83.0% respectively) than the positive control (17.0%) in E. frenatum. All other parameters did not vary significantly among treatments. These results indicate cGnRH IIa can be successfully used as a spawning aid in both species.     

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.     

Gonadal steroidogenesis and the possible role of steroid glucuronides as sex pheromones in two species of teleosts

In general, female zebrafish,Brachydanio rerio, ovulate only in the presence of males. The stimulant must be pheromonal as even male holding water is capable of inducing ovulation. After ovulation the mating phase begins. During this phase the male follows the female and oviposition as well as fertilization takes place. Both the ovulation and the mating are controlled by pheromones synthesized by the gonads. Ovulation can be induced by testicular homogenates. After the lipid material has been extracted from the testicular homogenates, the remaining aqueous phase can still induce ovulation. However, when the aqueous phase is treated with the enzyme-glucuronidase, it loses the ability to induce ovulation. This is an indication that glucuronides, probably steroid glucuronides, are the compounds responsible.During the mating phase, ovulated female zebrafish become attractive to males. It was found that, after ovulation, ovarian extracts contain the compounds responsible for attracting males. The attractant consists of a mixture of steroid glucuronides.After incubation of the gonads with³H-precursors seven steroid glucuronides have been identified in the testis and five in the ovary.Under fish culture conditions the African catfish,Clarias gariepinus, can produce postivitellogenic oocytes throughout the year. However, in capitivity neither males nor females spawn. In female catfish maturation and ovulation can be induced by treatment with gonadotropins. It might be possible that, analogous to the zebrafish, some reproductive processes in the catfish have to be induced by pheromones. It has been demonstrated that pheromonal compounds released by the seminal vesicles are involved in the attraction of female conspecifics. The steroid glucuronide synthesizing capability of the testes and the seminal vesicles of the male catfish are examined, as well as that of the ovary before and after ovulation of the female catfish. Both testes and seminal vesicles appear to be capable of steroid biosynthesis but only the latter synthesizes steroid glucuronides. Six of these conjugates have been isolated and identified. In the female catfish the ovaries are capable of synthesizing seven steroid glucuronides, but only after ovulation.     

Seasonal reproductive cycle of Waigieu seaperch (Psammoperca waigiensis)

Predictive and reliable parameters of reproductive status are integral aspects of sustainable fisheries and aquaculture management. These parameters are also important for an accurate evaluation of the effects of different treatments on sexual maturation in fish farming. In the present study, we have characterized the seasonal reproduction profile and described changes in sex steroids in relation to gonadal maturation and development in female and male Waigieu seaperch (Psammoperca waigiensis). The experimental period covered a full calendar year (January–December). In males and females, we observed that plasma sex steroid hormones [oestradiol‐17β (E2), testosterone (T), 11‐ketotestosterone (11‐KT) and progesterone (P)] levels showed monthly fluctuations during the spawning period. Particularly, plasma steroid hormone levels were positively associated with gonadosomatic index values. In addition, high levels of plasma steroid coincided with recruitment of oocytes into yolk accumulation in females. The main spawning period occurred between April and October in females, and between March and November in males. The non‐aromatizable androgen, 11‐KT is generally believed to be the active male‐specific androgen in teleosts, and is associated with the process of spermiation, development of secondary sexual characteristics and regulation of male reproductive behaviour in most teleost species. In this study, we found relatively high amounts of 11‐KT in females between May and December, suggesting an integral role in the maturation process, also for the females. A rapid peak in plasma P level was observed in November and suggests significant roles during post‐spawning and/or resting periods in both female and male fish. Furthermore, all oocyte developmental stages were present within the same sampling month and also within the spawning period, demonstrating the gamete group asynchronous developmental strategy. Overall, Waigieu seaperch showed strong seasonality in reproductive development with corresponding sex steroid patterns. The data presented in this study may contribute to the understanding of the reproductive endocrinology of a tropical marine finfish with increasing industrial prospects and sustainable aquaculture of this species in a developing country, such as Vietnam.     

Induction of spawning in Pacific white shrimp Litopenaeus vannamei (Boone, 1931) by injection of its molt inhibiting hormone isoform II produced in E. coli

The optimization of reproductive parameters in shrimp farming continues to be a challenge for most producing countries. Although the crustacean neuropeptides have been studied extensively in the last two decades, the functions of most of these neuropeptides remained putative. Among them, molt‐inhibiting hormone isoform II (MIH II) has shown an important role in vitellogenesis. In this study, the cDNA encoding mature MIH II peptide was isolated by RT‐PCR from the L. vannamei eyestalk. The cDNA was cloned into pET28a bacterial expression vector. Recombinant MIH II was obtained in the form of insoluble inclusion bodies and purified to ~88% purity. Two doses of rMIH II and a negative control group were assayed in vivo. The stages of ovarian maturation and spawning were recorded during 72 hr post‐injection. The results showed that ovarian maturation occurred approximately in 9% and 33% of females injected with rMIH II at the doses of 300 and 600 ng/gbw respectively. Neither maturation nor spawning was detected in the negative control group. Females injected with 600 ng/gbw, which showed vitellogenic stages III and IV, spawned. These preliminary results argue that the hormone rMIH II could be a promising candidate to induce spawning in L. vannamei shrimp.     

Induced maturation and spawning of milkfish,Chanos chanos Forssakal,by hormone implantation

The milkfish, Chanos chanos Forsskal, does not reach gonadal maturity easily in captivity. In an attempt to induce maturation, exogenous hormones, LHRH-A and 17α-methyl-testosterone, were implanted into adult milkfish either alone or in combination. The hormones were delivered using cholesterol pellets (LHRH-A) or silastic tubing sealed with elastomer (17α-methyl-testosterone). The fish were implanted three times at monthly intervals between March and May of 1985. The combination of LHRH-A and 17α-methyl-testosterone induced significantly more maturing fish (P < 0.05) than LHRH-A alone or sham controls; 88%, 38%, and 13%, respectively.Fish with average egg diameters between 768 μm and 905 μm, spawned 48 h after hormone implantation. These results indicate that the maturation and spawning of milkfish in tanks can be induced and accelerated 1–2 months earlier than the beginning of the normal spawning season through hormone implantation.     

Induced Spawning of Striped Snakehead,Channa striatus,Using Ovatide

ABSTRACT

An experiment was investigated to evaluate the effects of different doses of a new spawning agent Ovatide on induced spawning of striped snakehead, Channa striatus. Nine matured females with swollen abdomen and 18 male fishes with body weights ranging from 750 to 900 g, were randomly assigned to three treatment groups. Fishes in each group were administered 0.2, 0.4, and 0.6 mL of Ovatide/kg body weight, respectively. Each breeding set consists of two males and one female. Partial spawning response was observed when the fishes were administered with 0.2 mL/kg BW of Ovatide. Complete spawning was noticed in the medium (0.4 mL/kg BW) and higher (0.6 mL/kg BW) dose administered groups. The latency period and number of eggs spawned ranged from 23 to 27 hours and 1,764 to 7,096, respectively. No significant difference (P > 0.05) was noticed in the fertilization and hatching rate between the medium and higher doses of Ovatide tested. Thus, C. striatus could be successfully induced to spawn using a synthetic gonadotropin-releasing hormone with a dopamine antagonist at the dose of 0.4 mL/kg BW under captive conditions at 29 ± 1.5°C.     

Reproduction of striped bass, Morone saxatilis (Walbaum), broodstock: monitoring maturation and hormonal induction of spawning

Abstract Levels of gonadal steroid hormones were quantified in an adult striped bass, Morone saxatilis (Walbaum), broodstock during their gametogenic cycle. Blood plasma concentrations of Estradiol (E₂) and testosterone in females, or 11-ketotestosterone(11-KT) and testosterone (T) in males, were used as indicators of maturation. In both sexes, hormone levels were low in summer but increased significantly by late October to intermediate levels which were then maintained until late January. They then increased again rapidly to maximum pre-spawning values attained in late February or March, and subsequently decreased during the spawning period (April and May) with an increased incidence of spent fish with low hormone levels. The changes in blood hormone concentrations coincided with annual changes in photoperiod and water temperature that may be useful landmarks for maturation in captive broodstock. Mature females were implanted with pellets containing a dose of approximately 20 μg/kg body weight of [D-Ala⁶-Pro⁹-Net]-LHRH (GnRHa) in a matrix of cholesterol (CH) and cellulose. In April, they had not yet begun final oocyte maturation (FOM) and were too immature for conventional induction of spawning by injection with human chorionic gonadotropin (hCG). In early April, females given two 95% CH (slow hormone-release) GnRHa pellets (95/95) or females given one 80% CH (fast hormone-release) GnRHa pellet and one 95% CH GnRHa pellet (80/95) spawned within 13 days treatment (n= 4) with good egg fertility (76 ± 7% of total) and hatch rates (62 ± 15% of fertile). Females given dual fast-release GnRHa pellets (80/80) or control (Sham) pellets did not spawn or show evidence of increased oocyte diameter or development. In late April, four of six females given the 80/95 GnRHa pellet combination spawned within 9 days. Three fish produced fertile eggs (54 ± 18%). one spawned overripe eggs, and the remaining two increased oocyte diameter and maturation. Three corresponding controls did not spawn, and two of these showed clear signs of atresia within 11 days. In early May, some females were undergoing early FOM and were mature enough to be spawned by hCG injection. Three were given a single 80% CH GnRHa pellet and spawned within 6 days of treatment to produce fertile eggs (44 ± 6%). Of two other females given dual 80% CH GnRHa pellets, one spawned infertile eggs and the other failed to spawn within 9 days. GnRHa implants show promise as a technique for inducing spawning of captive striped bass broodstock although the optimum hormone delivery systems, dosages and release rates should be verified for fish at specific maturational stages.     

Evaluating spawning performance among captive Florida pompano Trachinotus carolinus broodstock using microsatellite‐based parentage assignment

Florida pompano has been identified as a promising candidate for commercial‐scale aquaculture production, but to date, little information is available regarding captive broodstock spawning characteristics. Genetic markers were tested for their power in monitoring mating outcomes and potential in analysing heritability of rapid growth trait in Trachinotus carolinus. A total of 20 unrelated adults (10 females and 10 males) were chosen for a hormone‐induced mass spawning event. The 515 fastest growing and 485 slowest growing fish of the total 4852 offspring were considered a selected progeny stock, and fish were collected at 45 days post hatch based on their growth traits. Parentage analyses based on the 20 breeders and 1,000 selected progeny were performed using a total of nine microsatellite markers, a 100% assignment rate was achieved, and a four‐marker set was the minimum number for the parentage assignment. The effective breeding number for the selected progeny was 11 (six females and five males), among which three females and two males were predominant contributors with the total contribution of 95.8% and 94.7% respectively. The proportion of fast‐growing offspring from broodfish and each mating cross (sire/dam) was used for detecting whether variation in growth of the offspring was related to parental stocks. Results showed that three adults and their mated combination exhibited the greatest fast‐growing offspring proportion (69.73% and 55.95%). This research provided new information regarding spawning performance and parental contribution during mass spawning events; both important first steps towards developing improved management strategies for captive Florida pompano broodstock.     

Reduction of early sexual maturation in male S0 Atlantic salmon (Salmo salar L.) by dietary supplementation of tetradecylthioacetic acid (TTA)

Effects of dietary tetradecylthioacetic fatty acid (TTA) on muscle fat, development of gonads and early sexual maturation in S0 Atlantic salmon during the first year in sea were investigated. TTA (0.5% w/w) was added to the feeds for 8 weeks in the spring. In May, at the end of the TTA‐feeding period, the fish in the TTA group had significantly (P < 0.05) less fat (10.1%) stored in muscle compared with the control group (10.8%). In September, mean male gonadosomatic index (GSI) in maturing fish in the TTA group was found to be lower compared with the maturing fish in the control group (P = 0.05). On the basis of GSI values, male sexual maturation in September was 10.0% vs. 14.4% for the TTA and the control group respectively. Thus, relative to the control group, the incidence of male sexual maturation in the TTA group was reduced by about 1/3 (P = 0.002). Production data was not affected by dietary supplementation of TTA. This study reveals that TTA significantly reduces the incidence of male sexual maturation in S0 Atlantic salmon. A significant elimination model of TTA in fish muscle that takes into account, the growth rate of the fish was further developed in this study.     

Studies on the Breeding of Pangasius sutchi Using Different Inducing Agents

A breeding experiment on Pangasius sutchi was conducted from July 2008 to September 2008 in a hatchery in West Bengal, India, using three different inducing agents, each injected at two different dosages: carp pituitary extract (CPE) and two synthetic hormones, Ovaprim and Ovatide. The fertilization rate was 82% to 91%. Hatching took place within 34 to 36 hours from the time of fertilization, with a hatching rate 73% to 79%. The response to spawning with CPE was best with Ovatide and Ovaprim in terms of percentage of fertilization, high hatching rate, and ease of stripping.     

Out-of-season spawning of pike perch (Sander lucioperca L.) without the need for hormonal treatments

The provision of fry and fingerlings, independent of the natural spawning season, can facilitate the implementation of innovative rearing strategies also in pike perch ( Sander lucioperca L.). As strict pharmaceutical acts or codes of conduct for organic aquaculture can constrain fish farmers in inducing spawning with hormonal applications, this study intended to develop protocols for advanced and postponed spawning just by simple photo-thermal treatments. After spending between 31 and 61 days at temperatures below 10 °C, different groups of pike perch spawners were treated with light and temperature programmes to advance spawning. Reproduction could be induced successfully 2 months before the natural spawning season when the mating pairs spent 43 or more days below 10 °C, followed by a maturation phase of 44–68 days at 15 °C and 16 h illumination per day. Advanced spawning could be documented for 32 out of 35 females (91%) that underwent photo-thermal treatments. Mean commercial fecundities up to 24% and average rates of developing eggs of 65% were observed in advanced spawning groups. Coldbanking of mature females allowed to postpone spawnings for 2 and 3 months. However, no egg development could be recorded in these treatment groups.     

Comparison of the lipid properties of healthy and pansteatitis‐affected African sharptooth catfish,Clarias gariepinus (Burchell), and the role of diet in pansteatitis outbreaks in the Olifants River in the Kruger National Park,South Africa

Pansteatitis has been identified in wild populations of sharptooth catfish, Clarias gariepinus (Burchell), and Nile crocodiles, Crocodylus niloticus Laurenti, inhabiting the same waters in the Olifants River Gorge in the Kruger National Park, South Africa. Mesenteric and pectoral fat tissue was investigated microscopically and by fatty acid analysis in healthy and pansteatitis‐affected catfish from both captive and wild populations. Variation in fatty acid composition between pectoral and mesenteric fat was noted. Composition of mesenteric fat differed between fish from various localities as a result of differences in diet. Pansteatitis in the captive population, resulting from ingestion of high amounts of dietary oxidized fat, reflected higher levels of unsaturated fatty acids within the mesenteric fat. Mesenteric fat of pansteatitis‐affected wild catfish was characterized by an increase in moisture content, a decrease in fat content and a decrease in stearic and linoleic acids. The n‐3 to n‐6 fatty acid ratio of mesenteric fat was higher in pansteatitis‐affected wild catfish than in healthy catfish from the same locality, reflecting higher polyunsaturated fat intake by pansteatitis‐affected fish. The possible role of alien, invasive, phytoplankton‐feeding silver carp, Hypophthalmichthys molitrix (Valenciennes), in the aetiology of pansteatitis in both catfish and crocodiles in the Olifants Gorge is discussed.     

Skipped spawning in female iteroparous fishes

It is often assumed that iteroparous fishes spawn annually once reaching sexual maturity, but this is not always the case. This paper reviews available information on skipped spawning in female teleost fishes. All instances of non‐annual spawning are described as one of three types (retaining, reabsorbing, resting), depending on where in the normal spawning cycle development has been interrupted. Retaining ripe eggs is caused by conditions experienced during the spawning season (fish density, mate availability, pollution), whereas failure to start vitellogenesis (resting) or the breakdown of all oocytes that enter into vitellogenesis (reabsorbing) is caused by factors experienced prior to the spawning season (primarily temperature and poor nutrition). It is speculated that the relative shortage of data on non‐annual spawning may be because of difficulties in identifying non‐reproductive individuals. In an attempt to rectify this situation, the criteria needed to identify females undergoing the three forms of spawning omission are presented in terms of external appearance of gonads, gonad indices, and histological analysis. The energy saved by not spawning in a poor year may lead to increased survival and the probability of spawning in subsequent years. As the cumulative number of progeny gained by surviving to spawn in multiple subsequent years outweighs the number of progeny lost by not spawning in a given single year, occasional omission of spawning may constitute an adaptive trait in long‐lived iteroparous fishes.     

An hypothesis on the light requirements for spawning penaeid shrimp,with emphasis on Penaeus setiferus

Using standard equations, we have concluded that the light intensities which have been used by investigators attempting to induce maturation and spawning are up to 4000 times higher than would occur over the spawning grounds of Penaeus setiferus, a species for which successful controlled spawning has yet to be achieved. We recommend that future attempts at captive maturation and spawning with that species be conducted at light levels of no more than 12 μW cm^?2 and that the intensity at which other species of shrimp are matured and spawned be examined relative to natural light levels on their spawning grounds. It is further recommended that the need for light of blue or green color be evaluated for its role as a promotor of maturation and spawning. The intent of these modifications in current spawning procedures is to eliminate the need for eyestalk ablation in captively spawned penaeid shrimp.     

Inducing oocyte maturation, ovulation and spawning in the African catfish, Heterobranchus longifilis Valenciennes (Pisces: Clariidae), using frog pituitary extract

Abstract. Oocyte maturation, ovulation and spawning were successfully achieved in the African catfish, Heterobranchus longifilis Valenciennes, using frog pituitary extract. During artificial spawning, oocyte maturation and ovulation were induced at a dosage of 7mg per kg of individual fish weight. Latency period varied from 7 h at 29°C to 11 h at 25°C. Dead eggs became opaque between 8 and 10h after fertilization while embryogenesis was found to be temperature dependent. Mean hatching and larval deformity rates were 63·08%± 7·56 and 8·08%± 1·93 respectively. During induced natural spawning, the extract was applied at a dosage of 35 mg per kg of female fish weight. Spawning occurred between 10 and 11 h after hypophysation. Mean hatching and larval deformity rates were 41·38%± 8·48 and 10·00%± 2·00 respectively.     

Immunoreactive somatolactin cells in the pituitary of young,migrating, spawning and spent chinook salmon,Oncorhynchus tshawytscha

Immunocytochemical techniques using an antiserum to cod somatolactin (SL) demonstrated the presence of SL cells in the intermediate lobe of the pituitary in Oncorhynchus tshawytscha. The cells were small in yearling fish. Two groups of maturing fish were studied. In the spring run salmon collected in April and May during the upstream migration, the SL cells appeared stimulated. In September, during spawning, SL cell stimulation was maximal with indices of hypertrophy and degranulation often more marked in females than in males. In the other group, salmon of the fall run collected in the Pacific Ocean in August had well developed gonads, large gonadotropes and abundant SL cells. In spawning salmon (September) the SL cells were stimulated, mainly in females. However, the final stimulation was less intense than in spring run spawning fish. The SL cells were smaller, without evident granule release, but still abundant in spent salmon of the fall run caught at the end of November. Various factors (time spent in rivers before spawning, starvation, decalcification, stress, hypothalamic influences) were considered which might explain differences between spring and fall run salmon. These observations suggest that SL may play a role in the control of gonadal maturation in chinook salmon as it may also do in sockeye and chum salmon previously studied, and that SL cells may be sensitive to the ambient salinity.