Use of endoscopy for gender and ovarian stage determinations in Russian sturgeon (Acipenser gueldenstaedtii) grown in aquaculture

The Russian sturgeon (Acipenser gueldenstaedtii) is native to the Caspian Sea, the Black Sea and the Azov Sea. It used to breed in the main incoming rivers, until dam construction in the mid 20th century blocked upriver spawning migration. Aquaculture of Russian sturgeon has only recently begun, prompted by their declining populations in natural habitats and the rise in meat and caviar prices. However, information on their gonadal development and puberty under culture conditions is incomplete.

Because sturgeons have no external sexual dimorphism and there are no external markers for sexing, internal examination of the gonads must be employed for gender identification as well as to monitor their development. The present study describes endoscopic and histological observations of the gonads of young Russian sturgeons aimed at identifying gender and monitoring ovarian developmental stage in females up to the age of 6 years, when they enter their first puberty cycle, as well as at 7 years of age, when they have completed vitellogenesis, under culture conditions. This information, as related to fish age and size, is of vital importance to commercial farming of Russian sturgeon for caviar production and reproduction.

For gonadal observations in both sexes, we used an endoscopic system consisting of a 4 mm, 14 cm long cystoscope sheath incorporated with fiber-optic light transmission, connected to a halogen cold light source and a miniature videocamera with a control unit attached to a color monitor. This system allowed us viewing of the fish's abdominal organs, and to save pictures of selected areas of the gonads on a computer as the fish's personal record. Ovarian biopsies were taken in parallel for histology at typical stages of gonadal development.

Gender could be identified with this system as early as at 3 years of age and the sex ratio under culture conditions of females, males and unidentified gender were 55, 40 and 5%, respectively.

Not only did large differences occur in the developmental stages of female of the same age group, but also ovarian development was highly asynchronous at the early vitellogenic stages. In late vitellogenesis, at the “gray egg” stage (1600–2600 μm diameter), the oocytes were quite regular in size and color, and remained so until the final stages of maturity.

Our study suggests that endoscopy is an efficient method for both gender identification at an early age, and for determination of gonadal development stage in sturgeon aquaculture. The ability to see the whole intact gonads of anesthetized fish can reveal important management and research information, with minimal damage or stress to the fish.     

Fluctuations in gonadotropin and ovarian steroids during the annual cycle and spawning of the common carp

The main objective of the paper is to describe the annual changes in hormones associated with reproduction in the female carp under the conditions prevailing in the Israeli fish culture. Fish were sampled monthly throughout 1984; gonadosomatic index (GSI) was calculated and the diameter of ovarian follicles was measured. Gonadotropin (GTH) content in the pituitary and the circulating GTH, estradiol, testosterone and 17, 20-dihydroxy-4-pregnen-3-one (17, 20-P) were determined by specific radioimmunoassays. GTH, estradiol and testosterone showed a bimodal annual pattern. The late summer peak was associated with initial vitellogenesis while the peak in spring occurred just before spawning, which took place in April-May. A resting phase in ovarian activity was noted in June and July. The levels of 17, 20-P were very low compared with those occurring during spawning induction. The paper summarizes a previous study by our laboratory on the changes in circulating hormones, as related to oocyte stages, in female carp induced to spawn by a GTH-calibrated pituitary extract. This study associates the short but prominent peak in 17, 20-P with the presence of follicles with maturing oocytes in the ovary. A correlation was found between the percentage of oocytes with eccentric germinal vesicle initially present in ovarian biopsies of females carp and their spawning success after hypophysation. The paper describes simple means to ensure successful induction of spawning in carp by utilizing a calibrated pituitary extract and prior selection of females that would respond to the induction treatment.     

Induced spawning of maturing milkfish (Chanos chanos Forsskal) with gonadotropin-releasing hormone (GnRH) analogues administered in various ways

The response of mature female captive milkfish to mammalian and salmon gonadotropin-releasing hormone analogues (mGnRH-A and sGnRH-A) was investigated. Prior to spawning, six groups of three females received (1) 10–16 μg mGnRH-A from an osmotic pump implanted intraperitoneally (IP); (2) 100 μg mGnRH-A from a cholesterol/cellulose pellet implanted IP; (3) 10 μg/kg mGnRH-A as an intramuscular (IM) injection; (4) 10–16 μg sGnRH-A from an osmotic pump implanted IP; (5) 100 μg sGnRH-A from a cholesterol/cellulose pellet implanted IP, and (6) a cholesterol/cellulose pellet without analogue implanted IP.

The most effective treatment was 100 μg sGnRH-A/fish given in a cholesterol/cellulose pellet; all (3/3) of the fish spawned. However, mGnRH-A was more effective (2/3) compared with sGnRH-A (1/3) if osmotic pumps were used to administer GnRH-A. If the dose and method of administration were not considered, then the salmon and mammalian GnRH analogues were equally effective (62–67%) for induction of ovulation and natural spawning in milkfish. Gonads of control fish regressed.

At the doses tested, injections or pellet implantations were more effective compared with osmotic pumps. All pellet-implanted and injected females responded to treatment and 75% (6/8) spawned; half (3/6) of the pump-implanted females spawned. Spawning occurred from 18 to 36 h after treatment.     

Long photoperiod delayed spawning and increased somatic growth in gilthead seabream (Sparus aurata)

The effects of extended photoperiods, mimicking the longest day of the year, were studied in 1- and 2-year seabream. The photoperiod regimes started in late July, 36 and 39 days after the summer solstice and continued for 11 months, well beyond the natural reproductive season of December–March. Regime 1 (long day, 15.5L:8.5D), which used natural and fluorescent light, reduced the incidence of maturity in both year classes and females did not spawn although some gonadal development was observed. Among all 1-year sampled fish of regime 1, a maximum of 5% became spermiating males (March) and 5% reached the yolk granule stage of vitellogenesis (VO3; 250–400 μm diameter) by May. Among 2-year sampled fish of regime 1, 45% became spermiating males and 25% were females, which reached the advanced vitellogenesis stage (VO4; 400–600 μm) by April. Regime 2 (skeleton photoperiod), consisting of natural light and a 1.5-h pulse of fluorescent light during the period 14–15.5 h after sunrise, postponed gonadal development and spawning for up to 3 months. In this regime, a maximum of 80% of 1-year sampled fish were spermiating males in February and a maximum of 10% were VO3 stage females in March. In the sampled 2-year fish, the maximum levels were 50% spermiating males in February and 25% VO3 stage females in March. Control fish, which were exposed to the natural photoperiod (29°34′N), spawned during their natural season. The maximum levels for 1-year sampled control fish were 95% spermiating males and no females in December, while among 2-year sampled fish, maxima of 75% males in February and 45% VO4 stage females in November. Final average weights of photoperiod treated fish (1-year=430 g—regime 1, 400 g—regime 2; 2-year=582 g—regime 1, 518 g—regime 2) were significantly greater (p<0.05) than control fish (1-year=341 g; 2-year=476 g). Daily feed consumption (g/100 g fish) dropped from an average of 1.83 to 0.93 g for 1-year fish during August–December and from 0.88 to 0.54 g for 2-year fish during the same period. This was correlated with reduced autumn and winter water temperatures (26–20°C summer to winter change) and increased fish weight in all treatments. Daily feed consumption was also affected by the onset of spawning in the control and regime 2 (skeleton photoperiod) treatments of both 1- and 2-year fish. The efficiency of feed utilization (FCR) and protein and energy retention were all affected by the photoperiod regimes and followed the same pattern of decrease as weight gain, regime 1 (long day)>regime 2 (skeleton photoperiod)>control.     

The use of luteinizing hormone releasing hormone analogue for ovulation induction in black sea bass (Centropristis striata)

Interest in commercial production of black sea bass has increased in recent years, but reliable spawning methods remain problematic. The objective of this study was to evaluate the effects of oocyte size and luteinizing hormone releasing hormone analogue (LHRHa) dosage and delivery systems on ovulatory success for in vitro fertilization. Vitellogenic females with maximum oocyte diameters of 400–625 μm were implanted with a 95% cholesterol–5% cellulose pellet containing 50 μg of LHRHa. Fish with maximum oocyte diameters < 450 μm failed to ovulate. In contrast, 90% of fish with 500 μm oocytes spawned within 36 h and 40% of this group ovulated a second time. All of the females containing oocytes > 550 μm ovulated. In a second experiment, females with uniformly vitellogenic oocytes (> 500 μm) and implanted with 50 μg LHRHa ovulated substantial numbers of eggs (45,000–192,000 eggs/kg body weight (BW), but fertility was consistently low (0–15%). In a third experiment, 19 of 39 females receiving implants containing 6.3–23.6 μg LHRHa/kg BW during the spawning season ovulated, but fecundity (17,000–339,000 eggs/kg) and fertilization (0–98%) were highly variable. When fish were grouped by developmental index, calculated as the number of oocytes with diameters > 400 μm/total number of oocytes measured, there were no statistical differences among groups with respect to the number of spawns, fecundity or fertilization success. In a fourth experiment, 11 of 13 females with a clutch of fully vitellogenic oocytes that were injected with 20 or 100 μg/kg BW LHRHa ovulated between 1 and 2 times on consecutive days. Five of seven given an implant containing 12.5 μg LHRHa ovulated one or more times. Fish implanted with shams or injected with vehicle alone did not ovulate in any of the experiments. No differences were found in the number of spawns, fecundity or fertilization success from fish receiving different doses of injected or implanted LHRHa. Incubation of pooled eggs produced 155,000 larvae (60% hatch) and 95,000 one-gram juveniles. These results demonstrate that injected or implanted LHRHa is effective for inducing ovulation in black sea bass with maximum oocyte diameters > 500 μm.     

Preliminary study on performance of triploid Thai silver barb, Puntius gonionotus

The triploid chromosome condition was induced in Thai silver barb (Puntius gonionotus) by application of cold shock (2°C) to eggs at time intervals after activation of 0.5 min with a duration of 10 min which resulted in mean triploidy yield of 72.5% at 9 months of age. Growth rate of the 2–9-month-old, cold shock group (0.1–6.2 g/month) did not differ from that of the control (0.1–5.7 g/month). Gonadal somatic indices (GSI) of presumed triploid males and females were lower than that of control (GSI values of the presumed triploids were 35.0–60.2% and 28.7–75.9% of control males and females, respectively). Spermatogenesis and oogenesis were retarded in triploids. However, all stages of spermatogenic cells were observed in triploid males, including few spermatozoa. Oocytes of triploid females did not undergo vitellogenesis while normal oogenesis was observed in diploids. Nuclear volume of red blood cells (RBCs) of triploid fish was 1.63 times larger than that of diploids.     

Impact of long‐term dietary exposure to lead on some reproductive parameters of a female Common carp (Cyprinus carpio L.)

During a long‐term 3‐year dietary exposure of mature carp females to lead, its bioaccumulation in the brain, changes in the neurohormonal activity at the level of the hypothalamus and the pituitary gland as well as the growth and maturation of ovaries were analysed. Moreover, an analysis of the effectiveness of hormone‐stimulated spawning during two subsequent spawning seasons was carried out. The results of the analyses show that chronic exposure of maturing and mature carp females to lead in feed results in its accumulation in the brain (1.365 μg g^?1). This impairs the endocrine activity of the hypothalamus, which is manifested by, among others, an increased secretion of dopamine and impaired spontaneous secretion of luteinizing hormone (LH). The observed decrease in the concentration of gonadotropin in blood inhibits vitellogenesis, which is manifested by a lower degree of the maturity of oocytes, lower fecundity (as measured by egg number, egg weight, egg maturity and egg survival) and lower gonadosomatic index values. The final negative impact of lead is the impairment of reproductive functions, as manifested by a smaller number of spawning females and in their lower fecundity during spawning.     

Enzyme-linked immunosorbent assay (ELISA) measurement of vitellogenin in plasma and liver histopathology in barfin plaice <Emphasis Type="Italic">Liopsetta pinnifasciata</Emphasis> from Amursky Bay,Sea of Japan

Vitellogenin (Vg) of the barfin plaice Liopsetta pinnifasciata was isolated and purified. In native polyacrylamide gel electrophoresis, Vg appeared as one band. After being subjected to sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS–PAGE), Vg fraction produced several polypeptides with molecular masses of 180, 98, 70, 52, 41 and 37 kDa. MALDI–TOF mass spectrometry (MS) of the 180- and 98-kDa Vg polypeptides from the SDS–PAGE gel and de novo sequencing of their four peptide fragments based on MS/MS analysis confirmed that the purified proteins were vitellogenins, which shared high similarity with the Vgs of the barfin flounder Verasper moseri and Atlantic halibut Hippoglossus hippoglossus. The most part of the predicted sequences obtained from the L. pinnifasciata 180-kDa polypeptide has previously been found in the V. moseri vitellogenin type B, the sequences obtained from the 98-kDa polypeptide were found in V. moseri vitellogenin type A, so these findings allow us to propose that L. pinnifasciata has at least two different forms of Vg. Rabbit polyclonal antibodies against Vg were produced, and a quantitative enzyme-linked immunosorbent assay was developed. The concentration of Vg in barfin plaice from the moderately contaminated area of Amursky Bay in the Sea of Japan was detected based on the maturity stage of their gonads. In November 2008, the Vg concentration in the plasma of females with advanced oogenesis varied from 5.295 to 28.367 mg/ml (mean 16.38 ± 6.73 mg/ml, CV = 41.1%); in the plasma of males, the concentration ranged from non-detectable to 0.957 mg/ml (0.29 ± 0.42 mg/ml, CV = 127.9%). In October 2009, the Vg concentration in female plasma was lower than in November 2008 (2.21–13.87 mg/ml). High individual variability of plasma Vg was characteristic for maturing males (CV = 200.3%) and immature females (CV = 255.5%), and there was no significant difference between plasma Vg concentrations in males captured in November 2008 and October 2009 or in maturing males and immature females. Vacuolisation of hepatocytes was more typical for males with low plasma Vg concentrations and females with high plasma Vg concentrations. Necrosis and pyknosis of hepatocyte nuclei were more frequent in males with high Vg concentrations and in females with low plasma Vg concentrations.     

Vitellogenesis with special emphasis on Indian fishes

Oocyte growth in most oviparous vertebrates including fish is due to the formation of yolk, and eggshell proteins (zona radiata proteins). Zonagenesis leads to the formation of zona radiata proteins in oocytes, which play an important role during oogenesis, whereas vitellogenesis leads to the formation of yolk in oocytes through a series of events during which the yolk precursor protein vitellogenin (Vg) is synthesized and secreted from liver into blood from where it is sequestered into the developing oocytes and thereafter proteolytically cleaved to form yolk proteins (YPs) and finally deposited in the ooplasm. Much research has been done in many fish species with respect to the number and nature of Vg and YPs and their probable functions during fish reproduction. Recent findings of multiplicity of Vg molecules in fishes reject the earlier view of a single-Vg model and have led scientists to explore the functions of individual Vg and their YP derivatives, lipovitellin, phosvitin, and β′-component. Two distinct types of Vg or Vg genes, containing or encoding the three YPs, have been detected in many teleosts. A third unusual, incomplete, phosvitin-poor Vg has been described recently in many fishes. In comparison to much of the information on vitellogenesis in many fishes very little is known for Indian fishes. In India research has been done in a few species such as the catfish, Heteropneustes fossilis and Clarias batrachus, the murrel, Channa punctatus and the Indian major carps, Labeo rohita and Cirrhinus mrigala. Immunological and biochemical analyses suggest the occurrence of multiple forms of Vg and their YP derivatives. The synthesis and incorporation of Vg are regulated by gonadotropin (GTH) and estradiol-17β (E₂). A differential role between estrone (E₁₎ and estriol (E₃) has been demonstrated for Vg synthesis. Enzyme-linked immunosorbent assays (ELISAs) for Vg have been developed to measure plasma Vg. Finally the different roles of Vg1 (HAI) and Vg2 (HAII) on vitellogenesis have been demonstrated. However, more research remains to be carried out in other fish species with respect to the number and nature of Vg and YPs and their genes in order to describe their reproductive functions.     

Induction of fertilizable eggs by conspecific vitellogenin implantation in captive female walking catfish,Clarias batrachus (Linn.)

The synthesis of vitellogenin (Vg) is induced by conspecific Vg (Vg1 and Vg2) and estradiol‐17β (E₂) as demonstrated by the pattern of ³H‐serine incorporation in the liver and plasma proteins. The incorporation studies indicated that the label was first incorporated into the liver after which it appeared in the blood in both E₂‐ and Vg‐treated male catfish. Since Vg was capable of inducing its own synthesis, experiments were conducted in females during preparatory–prespawning period (March–May) to make them gravid by implanting Vg pellets. Two implantations of 4 mg Vg1 pellets into female catfish with an interval of 15 days, followed by laboratory maintenance for 45 days of initial implantation showed a significant increment in ovarian weight with concomitant formation of yolky oocytes through synthesis and incorporation of Vg, whereas Vg2 implantation was not effective in this regard. Histological observation of yolky oocytes in Vg1‐treated group showed the peripheral migration of germinal vesicle (eccentric germinal vesicle), which indicates the onset of maturation. On 45th day, third implantation with 2 mg Vg pellets was performed and after 15 days, fish were hormonally induced with a single injection of hCG (2,000 IU/kg fish). Six groups were considered such as initial control, BSA‐implanted control, Vg1‐implanted, Vg2‐implanted, catfish collected from the field on the last day of the experiment and catfish collected during spawning period in this experiment with 3–7 fish in each group. Each of the experimental fish was sexually mature and the body weight was between 100 and 125 g. The percentage of ovulation and fertilization in the eggs of Vg1‐implanted group was 91% and 78%, respectively, which was almost similar to that of gravid female catfish collected during breeding period (July). The breeding performance in BSA‐ and Vg2‐treated females was very poor. The fertilized eggs were hatched in the laboratory conditions. Thus, in the female catfish, Vg1 not only induces vitellogenesis but also makes the oocytes viable for fertilization.     

A histological study of gametogenesis in captive red snapper Lutjanus campechanus

Gametogenesis was monitored histologically in wild‐caught red snapper (Lutjanus campechanus, Poey) maintained in captivity under simulated natural photothermal conditions. Gonads were collected every 2–3 weeks (average n = 14) for histology during the pre‐spawning season (February to May, temperature increasing from 16°C to 24°C). Primary vitellogenic oocytes were first observed in one female when temperature reached 20°C. Subsequent samples revealed females in pre‐vitellogenesis or at early stages of vitellogenesis, although one female had tertiary vitellogenic (Vtg3) oocytes. The first histological signs of spermatogenesis were observed when temperature reached 17°C. Spermatozoa were observed in testicular lobules of all males sampled on 14 May (24°C) but little or no sperm was released during manual stripping. Ten males and 10 females were left in tanks and monitored for spawning. No egg release was observed during the monitoring period that encompassed the natural spawning season of wild red snapper. Ovarian biopsies taken during the late spawning season (16 July) revealed that four of eight sampled females had Vtg3 oocytes. Males were manually stripped but released no sperm. These results indicate that captive red snapper can complete gametogenesis in photothermal controlled systems. Additional research is needed to develop procedures that will achieve reliable maturation and spawning.     

Shifting the natural spring–summer breeding season of the Australian freshwater crayfish Cherax quadricarinatus into the winter by environmental manipulations

Ninety sexually mature Cherax quadricarinatus females were exposed to various combinations of photoperiod and temperature for 2 months during the summer. Females were randomly assigned to either “winter” “semi-winter” or “summer” simulation treatments. In the “winter” treatment, crayfish were exposed to a simulated winter photoperiod (gradual decrease from 14L:10D to 10L:14D, 4 weeks at short day length followed by gradual increase to 14L:10D) and temperature (gradual decrease from 27 to 15 °C, held for 4 weeks, and then gradual increase to 27 °C). In the “semi-winter” treatment, crayfish were exposed to a simulated winter photoperiod and a summer temperature (27–29 °C). In the “summer” treatment, the crayfish were exposed to summer water temperatures and a photoperiod of 14L:10D. Following the 2 months of conditioning, the females were stocked for 7 months in small groups with males under environmental conditions similar to those of the “summer” treatment. All females were individually tagged and molting and spawning events were monitored. Females exposed to “semi-winter” conditioning in the summer, demonstrated a threefold increase in the rate of first spawning during the winter (December–February) compared with the other females. Crayfish breeders can easily implement these findings since shifting the breeding season into the winter only requires shortening of the photoperiod in the summer. The stocking of ponds in the spring with large nursed juveniles that hatched from eggs spawned in the winter, would allow the attainment of market size at the end of the limited growout season in temperate zones.     

Effects of Phase-Shifted Photoperiod Regimes on Oocyte Growth and Hormonal Profiles in Female Striped Bass Morone saxatilis

Abstract.— The use of 12–mo long, but phase-shifted advanced and delayed photoperiod cycles in the regulation of the reproductive cycle was investigated in captive-reared female striped bass Morone saxatilis during the 3-yr study in an attempt to control the timing of sexual maturation under simulated photoperiod conditions. Phase-shifted photoperiod cycles did not induce a full shift in oogenesis during the first year cycles, but did in the following years. Spawning time, indicated by maximum oocyte diameters, was advanced up to 4 mo in females maintained under the phase-shifted advanced photoperiod, and delayed up to 4 mo when they exposed to the phase-shifted delayed photoperiod, compared to the natural spawning time in Spring (March-May). Phase-shifted photoperiod regimes shifted the profiles of plasma testosterone (T) and estradiol (E₂), corresponding to the shift of oogenesis in the respective groups. Significant increases in T and E₂ levels occurred during the vitellogenic phase, and these levels peaked before the occurrence of maximum oocyte diameters. The studies demonstrate that phase-shifted photoperiod regimes can be used to control oogenesis, and have implications for ensuring the year-round supply of mature female striped bass, particularly in domesticated striped bass.     

The use of synthetic LH-RH analogue to spawn Chinese carps *

Abstract. The synthetic LH-RH analogue, des-GLY¹⁰[D-Ala⁶] LRH ethylamide was shown to be effective for induced spawning of Chinese carps. Ovulation was induced in 23 of 34 female grass carp, Ctenopharyngodon idella Val., receiving a single injection of synthetic LH-RH analogue. Spawning success was 92% for 3-year and older fish at optimum temperatures (22–26°C). Reduced spawning success was noted for 2-year-old female fish, and when temperature exceeded 27°C. No mortality of study fish was observed following a single injection. Hybrid carp, grass carp x bighead carp, Aristichthys nobilis Val., and grass carp x silver carp, Hypophthal-michthys molitrix Val., were produced using synthetic LH-RH analogue. Male grass carp, bighead carp and silver carp exhibiting pearl organs were stimulated to produce milt regardless of the presence or absence of free flowing milt at the time of injection. The potential importance of synthetic LH-RH analogue in fish culture and mechanisms of hormonal action are briefly discussed.     

Purification and development of ELISAs for two forms of vitellogenin in Indian walking catfish, <Emphasis Type="Italic">Clarias batrachus</Emphasis> (L.)

Two forms of vitellogenin (Vg: Vg1 and Vg2) were purified from the plasma of estradiol-17β (E₂)-treated Indian walking catfish, Clarias batrachus, by gel filtration and adsorption chromatography. Native Vg1 and Vg2 had apparent molecular masses of 375 and 450 kDa, respectively, and both Vgs resolved into two similar major bands (95 and 67 kDa) in SDS-PAGE under reducing condition. Polyclonal antisera raised against each form of Vg were absorbed with a combination of hypophysectomized male catfish serum proteins and alternate Vg to ensure specificity. Immunological analyses verified the presence of Vg1 and Vg2 in the plasma of female catfish. Homologous ELISAs were developed for Vg1 and Vg2 using their respective harvested antisera, which exhibited the detection limit of 100 ng ml^?1 for Vg1 and 40 ng ml^?1 for Vg2, and low level of cross-reactivity (not parallel to the standard) was found with alternate Vg in each assay. Treatment of male catfish with E₂ induced both Vgs showing a proportionate ratio of Vg1 to Vg2 at 5.6:1. Plasma concentrations of both Vgs measured by ELISAs at different reproductive phases of field collected female catfish increased in accordance with the ovarian development, keeping the proportionate ratio of Vg1 to Vg2 at about 2:1 in fish undergoing vitellogenesis during prespawning period and 1:20 during spawning period, suggesting that Vg1 may be the major Vg to contribute in yolk formation, whereas Vg2, besides its role in yolk formation, may facilitate other physiological functions. The present study, thus, demonstrates the occurrence of two unequally synthesized Vgs in the catfish.     

Off flavor characterization and origin in French trout farming

The results of a study on off-flavor problems in four French trout (Onchorynchus mykiss) farms are presented. Methodological aspects on sensory analyses and volatile compounds quantification in fish are discussed. Detection of odorous compounds by gas chromatography–mass spectrometry shows that significant concentrations of geosmin (up to 18 μg/kg in fat) are found in trout. The sensory evaluation, the presence/absence of geosmin in the flesh and the identification of off-flavor compounds producers demonstrate the implication of Microcoleus in the appearance of earthy/musty off-flavors. The presence of this particular cyanobacterium is linked to the deterioration of water quality during the water recirculation period. Correlations between chemical and sensory detection in the flesh indicate that the taste evaluation enables the differentiation of the four categories “non-tainted”, “slightly-tainted”, “tainted” and “strongly-tainted”. However, the average concentrations of geosmin found for these different intensities are relatively limited, from 0.2 to 4.9 μg/kg. Finally, recommendations are made to allow a more effective control of off-flavor occurrences.     

Oocyte Growth in the Striped Mullet Mugil cephalus L. Maturing at Different Salinities

Profiles of oocyte growth were obtained from female striped mullet Mugil cephalus L. held in salinities ranging from fresh water to seawater during two consecutive spawning seasons (1988–1990) in Hawaii. Females underwent vitellogenesis at all salinities (i.e., 32–35, 13–20, and 0%‰) tested. Females maturing in fresh water exhibited a slower rate of oocyte growth, and a significantly lower number completed vitellogenesis. All females were induced to spawn in full seawater. The number of fertilized eggs per spawning was highest from females maturing in brackish water. More females were able to be induced to spawn twice in brackish water during the 1989–1990 season than in fresh or seawater. The results from the current investigation suggest that salinities ranging from 13–35 ppt are adequate for ovarian maturation in captive striped mullet females.     

Induced spawning of hatchery-raised Brazilian catfish, cachara Pseudoplatystoma fasciatum (Linnaeus, 1766)

In the months of January 2001 and 2002, female cachara Pseudoplatystoma fasciatum were selected during their first and second gonadal maturation (2 years and 7 months old and 3 years and 7 months old, respectively) with an of oocyte diameter of 937.5 μm (82.5% with central nuclei and 17.5% with peripheral nuclei). Nine females in first maturation received two doses of carp pituitary extract (CPE), 0.5 mg/kg and 5.0 mg/kg; seven received two doses of human chorionic gonadotropin (hCG), 5 and 10 IU/g; five received doses of 0.5 CPE mg/kg and 5 hCG IU/g (CPE+hCG); and four received 0.9% saline (saline). Nine females from CPE and seven from hCG presented oocytes with the same diameter at the moment of oocyte release (100% with germinal vesicle breakdown and fertilization rate of 53.44±18.3 and 54.81±11.8%; larvae number of 165,330±94.1 and 158,570±20.6, respectively). The five females from CPE+hCG did not respond to the hormonal treatment. The four females from the saline group did not ovulate. In January 2002, 6 of 15 selected females that were going through the second reproductive cycle received CPE (five received hCG and four received saline), showing oocyte diameters similar to the ones in the first maturation. At stripping, CPE females had an oocyte diameter of 1062.5 μm (the hCG females had oocyte diameters ranging from 937.5 to 1125.0 μm; fertilization rates of 56.08±30.9 and 81.90±17.3%; 364,547±244 and 633,129±190, larvae, respectively). The fertilization rates and larvae number were higher in the second gonad maturation, both for CPE and hCG.     

Tank spawning of grass carp

Induced spawning of grass carp in indoor tanks without hand stripping of gametes is described. Circular tanks (1.83 m diameter) with biofiltration units were utilized for conditioning and spawning. This method was found to be successful and has several advantages over the dry-fertilization method: (1) preliminary ripening and spawning is conducted in small systems where the environment is controlled; (2) manpower requirements are reduced; (3) handling of brood fish is reduced, thereby minimizing mortalities; and (4) spawning occurs when the fish is physiologically ready, insuring good egg quality.     

Stimulation of ovarian growth by methyl farnesoate and eyestalk ablation in penaeoidean model shrimp,Sicyonia ingentis Burkenroad, 1938

The effect of methyl farnesoate (MF) administration on the vitellogenesis of the penaeoidean shrimp, Sicyonia ingentis, was studied. The short‐ and long‐term treatment effects as well as the effect of two MF injection regimens (0.1 and 1.0 μg MF/injection) were evaluated. The studies were also carried out to understand the pattern of vitellogenesis in eyestalk ablated adult and juvenile shrimps. A combination of endpoints, haemolymph vitellogenin (Vg) levels, gonadosomatic index (GSI) and histology, was used to study the effect of these treatments. The GSI increased in all the MF‐treated shrimp compared with the control shrimps. Although haemolymph Vg levels declined over the experimental period in all the treatments, the Vg levels decreased significantly only in the short‐term treatment with 1.0 μg MF. Similarly, haemolymph protein level also declined over the experimental period in all the treatment groups. However, except in the long‐term treatment with 0.1 μg MF, all treatments showed a significant decrease in haemolymph protein level. Conversely, in all eyestalk ablated adults and juveniles, haemolymph Vg, total protein and GSI increased over the experimental period, all of which were higher than the concurrent control. The discrepancy in the vitellogenic pattern between MF‐treated and eyestalk ablated shrimp was possibly due to the difference in the ovarian phase of the initial control. Although unilateral eyestalk ablation failed to induce vitellogenesis in juveniles, bilateral ablation induced vitellogenesis, which indicates that juveniles are competent to undergo vitellogenesis.