Sustained, Natural Spawning of Southern Flounder Paralichthys lethostigma Under an Extended Photothermal Regime

Hormone‐induced spawning of southern flounder Paralichthys lethostigma has produced substantial numbers of viable eggs, but wide variations in fertilization and hatch rates have been reported. Recently, sustained natural spawning of southern flounder broodstock, without hormone induction, has been achieved in our laboratory. Adults (average weight = 1.12 kg; N= 25), including 6 captured as juveniles in 1993 and 19 captured as adults during September 1998, were stocked in two 4.8‐m³ controlled‐environment tanks in October 1998 and held under natural photothermal conditions until January 1999, when an artificial winter photo‐period of 10 L:14 D was initiated and then maintained through April 1999. Sex ratio was approximately 13 females:8 males:7 unknown. Natural spawning was observed in early December 1998 and increased in frequency to a peak in March 1999, before declining in late April. Water temperature ranged from 13.9 to 24.5 C during the spawning period. Natural spawnings over 142 d produced a total of 18.3 × 10⁶ eggs, with a mean fertilization rate of 28.0% (range = 0–100%), yielding 4.94 × 10⁶ fertilized eggs. The mean percentage of eggs that remained buoyant in full‐strength seawater (34 ppt) was 41.3% (0–98%), while hatching rate of buoyant eggs was 37.3% (0–99%) and survival of yolksac larvae to the first‐feeding stage was 30.2% (0–100%). Gonadal biopsies in late April identified six females from both tanks as probable spawners. A preliminary comparison suggests that natural spawning produced much larger numbers of viable eggs per female, with higher egg quality (i.e., fertilization and hatching success) than hormone‐induced spawning. In contrast to natural spawning, hormone‐induced strip‐spawning enabled timing of spawnings to be more precisely controlled. These results suggest that a combination of both natural and hormone‐induced spawning of photothermally conditioned fish will help produce the large numbers of eggs required to support commercial production.     

Effects of photoperiod on growth and spawning efficiency of Nile tilapia (Oreochromis niloticus L.) broodstock in a recycling system

This study was conducted to evaluate the effects of photoperiod on the growth, feed conversion ratio (FCR) and spawning performance of Nile tilapia (Oreochromis niloticus) broodstock. Duplicate groups of two males (66±3.04 g) and six females (50.5±1.58 g) were stocked in 0.4 m³ fibreglass tanks in a recirculating water system at a male:female ratio of 1:3. The fish were subjected to four photoperiod treatments: 24:0 light:dark (L:D), 18L:6D, 12L:12D and 6L:18D. Light intensity was kept constant at about 2500 lx throughout the study. The fish were fed a commercial tilapia diet (35% crude protein and 16.6 MJ GE kg⁻¹) at a daily rate of 2–3% of tank biomass, twice a day, for 130 days. Males grew significantly faster than females under all photoperiod regimes. The best growth rate and FCR were attained at 18L:6D, followed by 24L:0D, 12L:12D and 6L:18D respectively. The number of eggs per female, number of eggs per spawn and number of spawnings per female were all significantly higher in the 12L:12D treatment than in all other photoperiod cycles. Interspawning intervals and days elapsed per spawn were also shorter in the 12L:12D treatment. The time to first spawning was slightly longer in the 24L:0D and 6L:18D than in 12L:12D and 18L:6D light phases. The 18L:6D and 6L:18D photoperiods produced the lowest spawning performance. It is concluded that a 12L:12D photoperiod regime should be adopted for maximum fecundity, seed production and spawning frequencies of Nile tilapia broodstock reared in intensive, recirculating systems. If maximum reproduction is desired, a near‐natural day length photoperiod should be used.     

Seasonal shift in spawning of Atlantic cod (Gadus morhua L.) by photoperiod manipulation: egg quality in relation to temperature and intensive larval rearing

Commercial intensive fry production of Atlantic cod will be dependent on production of viable eggs independent of season. This can only be done by manipulation of maturation by photoperiod, but little is known about potential effects on egg characteristics and larval viability. In two cod broodstocks, maturation was successfully advanced or delayed 6 months compared with normal spawning season (March–April) by manipulation of photoperiod. The advanced broodstock spawned both in spring and autumn the same year. In two of the spawning tanks during autumn, ambient temperature was reduced after reaching 13.7°C during the first half of the spawning period. Egg quality and viability were monitored, and several egg batches were incubated, hatched and start‐fed for examination of growth and survival. Temperatures above 9.6°C resulted in significant reductions in fertilization and normal egg development. Concurrently, fractions of dead and unfertilized eggs increased with elevated temperature. Actual relative fecundity was not affected by temperature. Egg characteristics improved when temperature was controlled and lowered below 9.6°C. Occurrence of irregular spawners suggests that handling of broodstock fish should be avoided during maturation and spawning. Cod larvae originating from eggs of the advanced or delayed broodstocks were successfully reared beyond metamorphosis. Survival was 9.0–46.6% and 29.3% in green and clear water respectively. Survival correlated with both initial and average feeding conditions, but growth rate did not correlate with either of survival and feeding conditions. Specific growth rates (8.3–13.6% day⁻¹) is comparable with other intensive rearing trials with cod, but were lower than reported from nature‐like systems.     

Environmentally conditioned,year‐round volitional spawning of cobia (Rachycentron canadum) in broodstock maturation systems

Year‐round control of the spawning cycle of cobia (Rachycentron canadum) has been established by using water temperature manipulation. To compare the effectiveness of using this method to induce volitional spawning in cobia, two 80 m³ recirculating aquaculture systems (RAS) were used. Temperatures in one of the maturation tanks (‘Mat 1’) were maintained between 27 and 29°C for 12 months of the 15.5‐month study period. Temperatures in the second maturation tank (‘Mat 2’) were allowed to fluctuate naturally throughout the year and ranged from 20 to 32°C. A total of 101 spawning events occurred in the tanks between the spring of 2008 and the summer of 2009 (3 April 2008 to 17 June 2009). Of the 38 total spawning events in Mat 1, 17 of them (44.7% of all Mat 1 spawning events) occurred during the off‐season (fall and winter). The egg viability rates did not differ significantly (P > 0.05) between on‐ and off‐season spawns in Mat 1. Conversely, cobia broodstock exposed to natural water temperatures (no environmental manipulation) in Mat 2 followed the natural pattern of warm water (>26°C) dependence, limiting egg production to spring and summer seasons. This method of water temperature manipulation allows for effective control of the cobia reproductive cycle without compromising egg viability.     

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.     

Spawning and Development of Larvae and Juveniles of the Rare Blue Mauritius Angelfish,Centropyge debelius (1988), in the Hatchery

This study documents the rearing and life history stages of the rare blue Mauritius angelfish, Centropyge debelius, from spawning of eggs through sexual maturity. A C. debelius pair was maintained at our facility for 22 mo. The pair was conditioned to spawn for a 4‐mo period in the fall of 2005 and a 4‐mo period in the summer of 2006 using water temperature and photoperiod manipulation. Continuous spawning was achieved at water temperatures between 22 and 24 C and a photoperiod of long day (LD) 13.5:10.5. Over a 124‐d period, the female produced 97 spawns, 59% of which resulted in fertile eggs. The average fecundity per spawn was 237 eggs (range 13–813 eggs). Fertility of all preserved spawns averaged 19.0 ± 19.8%. Larval rearing attempts using wild caught zooplankton and Artemia nauplii resulted in a total of 10 C. debelius juveniles raised through metamorphosis with an average larval survival up through metamorphosis of 0.36%. Three resulting juveniles were raised through sexual maturity. Sexual dichromatism was first observed about 180 d posthatch. A statistical model: Y = (X× 0.32437) ? (X²× 0.00043) ? 2.004, where Y = total length (mm) and X = number of days in culture, explained 97.6% of the variation in growth (P < 0.001, R² = 0.976). The development of eggs and larvae was observed to be similar to that of other Centropyge species that have been cultured. Results of this study indicate that the artificial propagation of C. debelius is technically feasible and forms the basis for this report.     

Seasonal differences in effect of broodstock diet on spawning success in the great scallop

Great scallop (Pecten maximus L.) broodstock collected at two different seasons, spring (May) and winter (December), were given four different diets during conditioning. The spring group was spawned in late June, and the winter group in February.The highest number of veligers in the summer spawning (30 millions per 10broodstock), was produced using a diet consisting of 80% Tahitian Isochrysis galbana. In the winter spawning, broodstock given a dietconsisting of 80% diatoms (Skeletonema costatum/Chaetocerosgracilis) produced the highest number of veligers (29 millions per 10broodstock). Fecundity and the number of egg-releasing individuals were higher in the early summer spawning (June) than in the winter spawning (February), however, the total number of three days old veligers produced was nearly the same in the two seasons.     

Dietary taurine improves reproductive performance of Nile tilapia (Oreochromis niloticus) broodstock

This study was conducted to evaluate the effects of supplemental taurine on reproductive performance of Nile tilapia (Oreochromis niloticus) broodstock fed soybean meal‐based diets. Four isonitrogenous (350 g kg^?1 protein), isocaloric (18 MJ kg^?1) diets were formulated to contain 0, 5, 10 and 15 g kg^?1 taurine. The diets were fed to triplicate groups of juvenile Nile tilapia (10–15 g average body weight) at a female: male ratio of 3 : 1, to apparent satiation, three times per day for 130 days. The size at first maturation decreased with increasing dietary taurine to 10 g kg^?1 and levelled off with further taurine supplementation. The time to first spawning was also significantly shorter at 10 g kg^?1 taurine level. Spawning performances, including spawning frequencies, total number of spawnings per tank, number of spawnings per female and absolute fecundity, were all significantly improved with increasing dietary taurine up to 10 g kg^?1. However, the quadratic regression analyses indicated that the maximum spawning performance occurred at 8 g kg^?1 of supplemental taurine. Eggs produced from broodstock fed 10 g kg^?1 taurine exhibited significantly higher hatchability and required shorter time for hatching and yolk‐sac absorption and also resulted in higher larval weight than at other dietary taurine levels. The highest egg protein, total amino acids and taurine were also obtained at 10 g kg^?1 taurine. These results suggest that 8 g kg^?1 dietary taurine is required for optimum reproductive outputs of Nile tilapia broodstock.     

Importance of broodstock holding temperature on fecundity and egg quality in three groups of photo‐manipulated Atlantic cod broodstock

We conducted a study to evaluate the effects of holding water temperature on the fecundity and egg quality of photo‐manipulated Atlantic cod broodstock. Adult cod Gadus morhua were distributed among several 25‐m³ tanks. Three separate photoperiods were used for each group to obtain three spawning per year (in May, July and December). While photoperiod was controlled, the temperature was not controlled and followed the ambient seasonal pattern. Due to the uncontrolled seasonal temperature used, each spawning group experienced different temperatures at a particular gonadal developmental stage. Otherwise, all groups were treated the same. Results showed that the May and July spawning groups released significantly more eggs per kg of fish (four and three times more respectively) than the December spawning group. The egg fertilization success and proportion of normal eggs were also higher in the May and July groups. Possible reason for the difference may be that the May and July groups experienced lower temperatures (4–7°C) during oocyte maturation and ovulation than the December group (7–9°C). Our results show the importance of using lower/natural temperatures during oocyte maturation and ovulation to obtain good quality eggs in photo‐manipulated cod broodstock.     

Biofloc technology in intensive broodstock farming of the pink shrimp Farfantepenaeus duorarum: spawning performance,biochemical composition and fatty acid profile of eggs

A 45‐day trial was performed to evaluate the effect of biofloc technology (BFT) with or without fresh food (FF) supplementation during pre‐maturation period on Farfantepenaeus duorarum spawning performance, biochemical composition and fatty acid profile of eggs as compared with conventional clear‐water system (CW+FF). Females raised in biofloc and that received FF supplementation (FLOC+FF) achieved better spawning performance in terms of number of eggs per spawn (49 × 10³), number of eggs per spawn per g of spawner's body weight (2.1 × 10³) and egg size (~275 μm) as compared with CW+FF (23 × 10³, 1.1 × 10³ and 263 μm respectively), but both treatments did not vary from FLOC (P > 0.05). High spawning activity was also observed in biofloc system as compared with clear‐water system as shown in number of spawns per ablated female (2.2–3.0 versus 0.6) and percentage of females that spawn at least once (80–82 versus 25%). Biochemical composition of eggs presented no significant differences among treatments. FA profile of eggs indicated that high spawning activity performed by females in FLOC+FF treatment was reflected in lower mean levels of EPA, DHA and sum of polyunsaturated fatty acids (n‐3) and (n‐6). The better reproductive performance demonstrated by females raised in biofloc justified the application of this technology in F. duorarum broodstock.     

Nauplii Production from Wild,Cultivated, and Mixed Populations of Blue Shrimp,Penaeus stylirostris

ABSTRACT

Due to the low nauplii production of cultivated broodstock, and to reduce the dependence on the wild stock, an experiment was carried out with 400 adult blue shrimp, Penaeus stylirostris, from wild and cultivated (F6) populations. Four treatments, each in duplicate, were applied: (1) wild females and males (W-W); (2) wild females and cultivated males (W-C); (3) cultivated females and wild males (C-W); and (4) cultivated females and males (C-C). More than 300 individual spawns were monitored to evaluate the egg and nauplii production per female. Mixed model ANOVA for factorial arrangements (4 × 3 × 2³ and 3 × 2³) were conducted. The factors considered besides the treatments were: rematuration (number of successive spawnings for a female), ovarian maturity, integrity of the spermatophore attached (complete spermatophore, “wings,” or “remnant”) and condition of spawning (partial or complete). The introduction of both wild females and males was a successful measure to improve the overall egg and nauplii production. Both mixed populations outperformed the cultivated broodstock, but were inferior to the wild stock (average production of eggs and nauplii: W-W-l 12,713 and 34,682, respectively; W-C-l 13,215 and 22,038, respectively; C-W-82,702 and 11,715, respectively; C-C-66,948 and 7,653, respectively). Populations with wild females produce a larger number of eggs, and wild males contribute to higher hatching rates. Other observations indicate the need to select for spawning only those females showing an advanced degree of ovarian maturity, having a complete spermatophore attached to the thelycum, and spawning completely.     

Additive effects of advanced temperature and photoperiod regimes and LHRHa injection on ovulation in Atlantic salmon (Salmo salar)

In order to evaluate manipulation of spawning time as a potential means to extend 0+ Atlantic salmon (Salmo salar) smolt production in Tasmania, Australia, female salmon were exposed to a natural/simulated natural (42°S) photoperiod or an advanced (L:D 9:15) photoperiod from the austral summer solstice (20 December) under natural or advanced (~ 6 °C below natural temperature) temperature conditions. In late summer (26 February) injections of a commercial LHRHa preparation or vehicle (propylene glycol) commenced. Regular ovulation checks were conducted and ova were fertilised using milt from LHRHa-injected males held under matching photo-thermal conditions. Plasma levels of 17β-estradiol (E₂) and testosterone (T) were monitored and reproductive success (cumulative % ovulation, % fertilisation and % survival to the eyed-egg stage) was recorded. Ovulations commenced first (09 March) in LHRHa-treated fish that experienced advanced photoperiod and thermal regimes whereas sham-treated fish exposed to natural photoperiod and temperature conditions where the last to ovulate (22 May-08 June). Treatment-related sequential changes in the timing of ovulations were reflected by sequential advances in the timing of peaks in plasma levels of E₂ and T. The fertilisation of ova from LHRHa-treated fish that experienced advanced photoperiod and thermal regimes was significantly reduced (~ 52%) relative to all other treatments (> 80%) but there were no significant treatment-related differences in the survivals of eggs to the eyed stage (~ 50-90%). Consequently, a maximum advance in the timing of median ovulation of 71 days and commercially acceptable eyed-egg yields were generated, demonstrating that combinations of photoperiod, thermal and hormone treatments may be employed to significantly extend spawning and thereafter increase the availability of 0+ smolts for grow-out.     

GnRHa‐induced Multiple Spawns and Volition Spawning of Captive Spotted Rose Snapper,Lutjanus guttatus,at Mazatlan,Mexico

Sexual maturation and induced spawning treatments were carried out with captive spotted rose snapper, Lutjanus guttatus. A total of 3013 × 10⁶ eggs (64.7% were floating) were produced from eight treated females in 42 spawns induced with GnRHa implants during the course of the present study. GnRHa ethylene‐vinyl acetate copolymer effective doses were 204 ± 11 µg/kg in June 2005, and 224 ± 13 µg/kg in July 2005. General fertilization was 50.9 ± 34.5% and 12–14 h after spawning, viability of floating eggs was 90.4 ± 12.4%. Mean incubation period at 29–31 C was 18–20 h, and mean hatching was 94.4 ± 8.2% (73–100%). Newly hatched larvae were 2.18 ± 0.15 mm in total length (TL). One month after the last hormone experiment, previously GnRHa‐treated and untreated fish began spawning voluntarily. Hormone‐treated breeders had higher fecundity than untreated fish, producing 72.5 million eggs versus 13.9 million eggs for the untreated fish, over the following 11 mo. Combined data of volitional spawning for total egg fertilization, viability, hatching, and larval TL were 77.7 ± 1.8%, 90.3 ± 1.3%, 87.9 ± 2%, and 2.50 ± 0.12 mm, respectively. These results can ensure the sustainability of a commercial hatchery.     

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.     

Effects of dietary highly unsaturated fatty acids and astaxanthin on the fecundity and lipid content of pond-reared Penaeus monodon (Fabricius) broodstock

Five diets that contained fresh squid meat as the basic constituent and were supplemented with different amounts of highly unsaturated fatty acids (HUFA) and astaxanthin were fed to pond‐reared Penaeus monodon broodstock. Diet A was sole squid meat. Diets B and C were supplemented with astaxanthin 50 and 100 mg kg^?1 respectively. Diets D and E were supplemented with HUFA 5 and 10 g kg^?1 and astaxanthin 50 mg kg^?1 respectively. The result showed that the group fed diet E had the best reproductive performance in all experimental groups. It had a higher proportion of spawns (71.5%), spawning rate (0.047), a shorter latency period (7.7±0.3 d), higher absolute fecundity (× 10³) (361.6±5.5) and egg production/female (× 10³) (597.0±18.0) than all the other experimental groups. The fatty acid composition in broodstock diets strongly affected the tissue and fecundity of broodstock. Good correlations between the content of 20:4n‐6 in eggs and the fecundity (r²=0.6109) and egg production (r²=0.9876) of broodstock were found. On the other hand, 22:6n‐3 and DHA/EPA ratio was negatively correlated with the fecundity of broodstock (r²=0.5362, 0.8702 respectively). The result also showed that the balance between n‐3 and n‐6 fatty acid families, total polyunsaturated fatty acids and total saturated fatty acid and 20:5n‐3 (EPA) and 22:6n‐3 (DHA) may play vital roles in maturation and reproductive performance of P. monodon broodstock.     

Multiple Induced Spawnings of the Indian Catfish Heteropneustes fossilis (Bloch) within a Prolonged Spawning Season

Abstract Singhi, the Indian catfish Heteropneustes fossilis , attained gonadal maturity repeatedly in a single spawning cycle by photothermal treatment. After the first spawning, maintenance at 30 C and 14 h light/10 h dark for 3 wk induced recruitment of the next batch of mature oocytes for subsequent spawnings. The number of eggs released declined with each spawning. However, fertilizability of the eggs was maintained until the fifth spawning. Mature fish were induced to spawn by D-Lys⁶ salmon gonadotropin-releasing hormone agonist administration. The dose of salmon gonadotropin-releasing hormone agonist for the second and subsequent spawnings was determined to be 25-fold less compared to the dose administered for the first spawning. Fish released the maximum number of eggs upon induction when they were maintained continuously at 30 C and 14 h light/10 h dark for 6 wk. The result obtained in the present study points to a possibility of obtaining multiple spawnings from those species of farmed fish which spawn once a year.     

Effect of Photoperiod Manipulation on Broodstock Spawning, Fertilization Success, and Egg Developmental Abnormalities in Atlantic Cod, Gadus morhua

Spawning activity in two captive Atlantic cod broodstock groups previously captured from the wild stock fishery was monitored over three spawning seasons. Both groups spawned under ambient photoperiod (PP) in the first year, after which photomanipulation was applied to compress the PP cycle of one broodstock group, while the second group was maintained on natural PP. Increased total egg volumes, number of egg batches, total egg production, and duration of the spawning season were observed in both ambient PP and advanced PP groups in the second and third spawning seasons. Compared to ambient controls, the photoadvanced group commenced spawning earlier, had a longer spawning season, produced a lower total volume of eggs over the entire spawning season, and had lower mean daily batch volumes of eggs, and the eggs were of smaller mean diameter. Fertilization success was >90% in all years, increased in both groups in the first year after photomanipulation but declined to prior levels in the following year. No consistent difference in fertilization success was observed between advanced PP and ambient PP groups. Rates of cell development abnormalities were low (<10%) in both groups over the three spawning seasons. Abnormalities of cell symmetry and clarity increased in both ambient PP and advanced PP groups over the three spawning seasons. However, no consistent differences in rate of cell abnormalities were observed between advanced PP and ambient PP groups within years. We conclude that PP advancement is an effective technique to compress the time between successive spawning periods in Atlantic cod and does not negatively affect fertilization success or rates of cell developmental abnormalities. However, it has a negative effect on both volume and size of eggs produced.     

Hatchery broodstock conditioning of the blue mussel <Emphasis Type="Italic">Mytilus edulis</Emphasis> (Linnaeus 1758). Part I. Impact of different micro-algae mixtures on broodstock performance

Blue mussel (Mytilus edulis) broodstock collected from the Irish Sea during wintertime (November) was conditioned with three different microalgae diets. Positive flagellates (PF) treatment consisted of Pavlova lutherii, Isochrysis galbana (T-Iso), and Chaetoceros calcitrans (1:1:1). Positive diatoms (PD) treatment consisted of Pavlova lutherii, Chaetoceros calcitrans, and Skeletonema costatum (1:1:1). Broodstock animals in the PF and PD treatments were fed a total of 2.4 × 10¹¹ algae cells per day. Animals in the negative flagellates (NF) treatment received only 1/8th of the total amount of algae of the PF diet. The conditioning diets had an impact on spawning success and broodstock fecundity but not on hatching rate, which was similar in all three groups. The best results were obtained with the PD diet where 84% of the conditioned animals spawned and females released 5.0 × 10⁶ eggs on average. Animals belonging to the PF and NF treatments released, on average, only 3.6 × 10⁶ and 1.6 × 10⁶ eggs, respectively. Although the amounts of algae provided to the broodstock animals had no effect on the hatching rate, the D-larvae resulting from the NF treatment were smaller in size than the larvae from the other treatments. Biochemical analysis of the different broodstock groups at the end of the experiment revealed higher carbohydrate levels in group NF than in PF and PD, supporting the theory that gametogenesis is mainly supported by the energy from the glycogen reserves. As far as we are aware this is the first study describing hatchery broodstock conditioning of blue mussels under fully controlled conditions.     

Oocyte and embryo quality in Crassostrea gigas (Portuguese strain) during a spawning period in Algarve,South Portugal

Egg quality is one of the most important factors determining larval viability. The study of oocytes released by `wild' broodstock will contribute to the definition of quality criteria applied to hatcheries as well as being an indicator of the oyster reproduction potential and aquaculture ongrowing systems. The aim of this study was to determine the relationship between condition of Crassostrea gigas spp. angulata (Portuguese strain) from `wild' broodstock (condition index, gonadal maturity and number of oocyte released), oocyte biochemistry and hatching rate of eggs during the spring/summer of 1995. Stages of gonadal maturity and condition index indicated that natural spawning of this species occurred between April and August with the highest spawning intensity occurring in June. Analyses demonstrated that C. gigas oocyte organic matter consist of 44–74 % proteins, 16–38 % lipids and 7–12 % carbohydrates. A significant relationship was observed between some biochemical parameters – organic matter and lipid content – and broodstock condition index and hatching rate. It seems that these two oocyte parameters (organic matter and total lipids) could be used to define quality of oocyte and thus larval viability. The oocyte protein content was also found to be associated with broodstock and larval quality. In summary, these data are a contribution to the knowledge of the reproductive cycle of oysters in Europe and may be useful in improving hatchery management of this species.     

Tolerance of Penaeus monodon Fabricius embryos to ozonated seawater

The tolerance of Penaeus monodon embryos from five spawnings (families) to four different ozone doses in seawater [0.0, 0.5, 1.0 and 2.0 mg L⁻¹, measured by the residual oxidant concentration (ROC)] was examined when applied for three exposure times (1, 2 and 4 min) at three post‐spawning treatment times (25, 120 and 480 min post‐spawning). Ozone dose typically had a larger affect on embryo hatching than exposure time and the ozone dose × exposure time interaction for most combinations of family and post‐spawning treatment time. At ozone doses of 2.0 mg L⁻¹, embryos had lower hatchings than controls for all families at 25 and 120 min post‐spawning, and for several combinations of family and exposure time at 480 min post‐spawning. At ozone doses of 1.0 mg L⁻¹, the effect on embryo hatching was more varied between families and exposure times for the three post‐spawning treatment times, but typically embryos were less affected when exposed at later post‐spawning treatment times. Ozone doses of 0.5 mg L⁻¹ typically had minimal effects on hatching for all exposure times and post‐spawning treatment times. In summary, later‐stage P. monodon embryos typically tolerated ozone doses of up to 1.0 mg L⁻¹ in seawater for durations of up to 4 min.