Oogenesis and relevant changes in egg quality of abalone Haliotis discus hannai during a single spawning season

Oogenesis of abalone Haliotis discus hannai was examined histologically during a single spawning season using broodstock of various maturation conditions, which were controlled by effective accumulative temperature (EAT). The quality of eggs spawned was determined in relation to oogenesis. For histological examinations, three to five females were sacrificed at 300, 600, 850, 1050, and 1150 °C days EAT, without induction of artificial spawning. Other females were successfully induced to spawn at 700 °C days EAT and were reared following spawning. Three of these females were then sacrificed every 200 °C days EAT until 1300 °C days EAT. Gonad histology showed that two oocyte cohorts matured in H. discus hannai ovaries during a single spawning season. One mature oocyte cohort could be spawned in multiple times. The second oocyte cohort started developing after the first oocyte cohort had been spawned or reabsorbed, and became fully mature 400 °C days EAT after the first cohort was depleted. For egg quality measurements, three to five females were successfully induced to spawn at 850, 1050, 1150, 1900, and 2350 °C days EAT (Experiment 1). Three females were induced to spawn twice, at 700 and 1500 °C days EAT, resulting in two batches of eggs from the same individuals (Experiment 2). Total lipid and protein content of eggs were measured and were greater in eggs from the second cohort than in eggs from the first cohort. No carbohydrates were detected in eggs and there was no difference in cytoplasm volume between the two cohorts. In hatcheries producing H. discus hannai, it is important to increase post-larval starvation tolerance by increasing the quality of eggs, to yield higher and more consistent survival. The results of this study suggest that H. discus hannai hatcheries should use eggs from the second oocyte cohort, which are of higher quality, rather than eggs from the first oocyte cohort.     

Life cycle of Caligus rogercresseyi, (Copepoda: Caligidae) parasite of Chilean reared salmonids

Caligus rogercresseyi, [Contrib. Zool. 69 (2000) 137] is the only caligid known to affect the salmon industry in Southern Chile. Economic losses due to reduced fish quality, cost of chemical treatment and outbreaks of other diseases such as the Piscirickettsiosis occur. The life cycle of C. rogercresseyi is described in rainbow trout reared in seawater tanks from observations made under natural conditions of light and temperature between January 1997 and April 1998. Fish were infected with laboratory-cultured larvae obtained from ovigerous females. Rainbow trout were periodically slaughtered for parasite collection and identification. C. rogercresseyi life cycle includes the following stages: two nauplius, one copepodid, four chalimus and the adult. No preadult stage was observed. Timing of the different stages of development was directly dependent on water temperature. The maturation of the eggs or the time for a complete life cycle took place at 45 days in July at 10.3 °C, 31–32 days in April at 12.4 and 12.8 °C, respectively, and at 26 days in November at 15.2 °C. In January, at 16.7 °C, only the appearance of first eggs were observed at 18 days. A simple degree–day (dd) model is proposed for each developmental stage between 4 and 17 °C, where the development rate is a linear function of the average temperature of water. Using this degree–day model, the proportion of fourth stage chalimus was maximum at 172 dd of effective temperature, adult males at 193 degree–days, adult females at 208 dd. The minimum temperature threshold is at 4.2 °C where there is no development of the parasite. The appearance of first eggs occurred at 231 dd and the first pigmented eggs at 277 dd. The temperature-independent degree–days value allowed to predict the timing of C. rogercresseyi life cycle at any temperature within the evaluated range.     

Eggs and hatchlings of the freshwater crayfish, marron (Cherax tenuimanus Smith), can be artificially incubated at high population densities, and are most successfully incubated at water temperatures between 20°C and 24°C

This study tested two premises. First, that there is a range in water temperature within which to artificially incubate marron eggs and hatchlings, which enables a combination of rapid development, high survival, and the production of large juveniles. Second, that the population density of marron eggs and hatchlings incubated artificially can be increased without altering development time, survival, or juvenile body weight. Marron eggs were collected from 31 gravid females and artificially incubated at four water temperatures (i.e. 16°C, 20°C, 24°C, 28°C), while eggs were collected from eight females and artificially incubated at three population densities (i.e. 0.35, 0.70, 2.80 eggs mL^?1). The eggs and hatchlings were assessed for development time and survival as they hatched and developed into juveniles. The juveniles were weighed at the conclusion of the incubation period. As water temperature was increased from 16°C to 28°C, there was an inverse curvilinear relationship between development time and water temperature, the time required for the eggs to hatch and develop into juveniles decreasing from 70.2 to 24.5 days. However, at 16°C and 28°C, the survival of the eggs and hatchlings was reduced (i.e. 83.8% and 87.7% at 20°C and 24°C vs. 70.4% and 57.5% at 16°C and 28°C), while incubation at 28°C resulted in the development of small juveniles (i.e. 29.7, 29.2, and 28.5 mg at 16°C, 20°C, and 24°C vs. 25.1 mg at 28°C). In comparison with water temperature, increasing the population density of the eggs and hatchlings from 0.35 to 2.80 eggs mL^?1 did not have a significant effect on development time, survival, or juvenile body weight. The two premises tested in this study were supported, demonstrating that marron eggs and hatchlings can be artificially incubated at high population densities, and are most successfully incubated in water maintained between 20°C and 24°C.     

Gametogenesis of the European flat oyster (Ostrea edulis) and Pacific oyster (Crassostrea gigas) in warm water in Israel

The gametogenic cycle of Ostrea edulis and Crassostrea gigas was studied in the outflow water of fish ponds in Eilat, Israel. Despite relatively high water temperatures (14–28°C) and high salinity (41 ± 2 ppt), both species developed gonads. O. edulis spawned from March to May, when water temperature ranged from 18 to 22°C, with a dormant period during the summer months. C. gigas spawned between May and August, when water temperature ranged between 20 and 26°C, with a dormant period between August and April. Spat never developed in the fish pond system.     

Effect of a sand substrate on the growth and survival of hatchlings of the softshell turtle, Trionyx sinensis Wiegmann

Softshell turtle (Trionyx sinensis Wiegmann) hatchlings were reared at 28 ± 2°C in the laboratory to test the effect of the presence of a sand substrate. The growth and survival of the hatchlings were monitored for a period of 14 weeks. The presence of the sand substrte significantly enhanced the growth of the hatchlings. Survival of the hatchlings, however, was independent of the presence of the sand substrate.     

Breeding and juvenile culture of the lined seahorse, Hippocampus erectus Perry, 1810

All seahorse species worldwide have been placed under CITES Appendix II since 2004, because they have been over-exploited for traditional Chinese medicine and aquarium trades. Aquaculture has been recognized as a long-term solution for sustaining the seahorse trade while minimizing wild collection. In this study, we evaluated the breeding and juvenile culture of an important aquarium seahorse species, the lined seahorse Hippocampus erectus, Perry 1810. Pairing, mating and copulation behavior were observed. Gestation time and brood size were 17.33 ± 2.94 days and 272.33 ± 66.45 individuals/brood, respectively. Growth rates differed among juveniles from different broods. Effects of temperature on the growth rates and survivorship of the juveniles during the first two weeks were compared. The highest growth rate and survivorship of the juveniles occurred at 28–29 °C among the temperatures tested (24–33 °C). Growth rate and survivorship of the juveniles during the first 9 weeks at 28 °C were investigated. The final standard length and survivorship of the juveniles were 6.32 ± 0.52 cm and 71.11 ± 10.18%, respectively, and the relationship between the wet weight and the standard length of the juvenile seahorses can be expressed as: W = 0.0034 L^2.5535 (r² = 0.9903, n = 12, P < 0.01). These findings suggest that H. erectus is a good candidate for commercial aquaculture.     

Mortality of yellowtail flounder, Limanda ferruginea (Storer), eggs: effects of temperature and hormone-induced ovulation

Yellowtail flounder, Limanda ferruginea (Storer, 1839), is a highly marketable fish species that used to be a significant groundfish in the northwest Atlantic region. The potential of yellowtail flounder for mass rearing has recently been investigated, but studies into aspects of mortality during early ontogeny are lacking. We investigated the impact of spawning inducement on mortality of eggs from fertilisation until hatch, and the effects of rearing temperature on mortality and hatching time in this multiple batch spawning flounder. An ash-free dry mass (AFDM) technique for determining mortality was employed. This technique reduced rearing tank disturbances and provided quantitative mortality estimates. We found (1) that spontaneously ovulated (SO) mean egg mass (AFDM 24.84±0.73 μg) was significantly higher than mean egg mass from induced ovulation (IO) (AFDM 20.09±0.54 μg); (2) that mortality of SO eggs was lower than IO eggs (p=0.028), (3) that SO eggs reached a relatively constant (maximum) mortality by about day 3 and IO eggs by about day 5; (4) that there was no significant difference in mortality between eggs incubated at 10 °C and those incubated at 4 °C (p=0.320); and (5) that time to 100% hatch took significantly longer for eggs incubated at 4 °C (256.0±73.5 h) than those incubated at 10 °C (77.9±11.6 h) (p=0.003). There was considerable variation in egg mortality among batches and females; nevertheless, general trends indicate that GnRHa-induction of ovulation tends to decrease egg quality. In addition, the AFDM technique was an easily employed and accurate method of determining mortality. We recommend that unless coordination of spawning is important to a particular rearing protocol, induced ovulation should not be employed.     

Effect of increasing salinity on physiological response in juvenile scallops Argopecten purpuratus at two rearing temperatures

Argopecten purpuratus can be cultivated using Recirculating Aquaculture Systems (RAS) as a method to increase production. In order to determine physiological response of A. purpuratus under different salinities and temperature conditions, two groups of juvenile scallops (small: h = 6.5 mm and large: h = 25.5 mm) were acclimated and close-cultured at salinities of 34, 38, and 42 g/l, at 16 and 22 °C and fed on Isochrysis galbana and Chaetoceros calcitrans. Survival, shell growth and scope for growth were determined at the end of the trials. Survival showed an inverse relationship with temperature and ammonia levels. In small scallops an increase in salinity at 16 °C increased survival. However, this relationship was not evident at 22 °C. On the other hand, salinity did not affect survival of large juveniles. Small juveniles had a lower survival (approximately 40%) than larger scallops (up to 85%) throughout the trials. Oxygen consumption was not affected by salinity. Small scallops showed similar oxygen consumption at 16 and 22 °C but in large juveniles higher values were registered at 22 °C. In large juveniles routine consumption at 16 °C was higher (up to 35%) than standard consumption. This pattern was not evident at 22 °C, suggesting that oxygen demand is higher regardless of feeding condition. NH₄⁺–N excretion rate is inversely related to salinity. Only small juveniles showed a higher NH₄⁺–N excretion rate at 22 °C. Scope for growth was positive in all treatments, although the upper limit of salinity should not be based only in this index. Higher scope for growth values at 38 and 42 g/l was related with a reduction in ammonia excretion and high absorption efficiency. In addition, an increase in salinity produced a reduction in NH₃–N proportion and under hypersaline conditions scallops tended to decrease excretion as a way of osmoconformation. This explains our findings of higher survival rates at higher salinities. Even though the scope for growth is positive at 42 g/l, the osmotic stress reduces the survival chances. The data obtained can be considered useful information for A. purpuratus culture under controlled conditions.     

Artificial incubation of Japanese crayfish (Cambaroides japonicus) eggs by using a simple, easy method with a microplate

We developed a simple, easy method with a microplate to artificially incubate Japanese crayfish (Cambaroides japonicus) eggs for their cultivation. We prepared 6-, 12-, 24-, 48-, and 96-well microplates containing sterile water heated to 5, 10, 15, and 20 °C. Fourteen experimental groups for each water temperature were prepared for each of different water volumes (0.125–10 ml) in each well. One embryonic egg was placed in each well. Experiments were also conducted with water collected from the lake where the eggs were harvested from and held at 15 °C. In the microplates with sterile water, high proportions of eggs hatched (60–100%) at 15 °C in all volumes of water, although the proportions of hatching were low (0–20%) at 5, 10, and 20 °C. All eggs died in the experiments that used lake water. We conclude that the 96-well would be the most effective size to hatch crayfish eggs in, because of its convenience. This method using a microplate is simpler and easier compared with methods of previous studies to artificially incubate crayfish eggs, and therefore it might be useful to incubate eggs of other freshwater crayfish species.     

Effect of Low-Temperature Incubation of Channel Catfish Ictalurus punctatus Eggs on Development, Survival, and Growth

To determine whether the embryonic period of channel catfish lctalurus punctatus could be extended at low temperatures, fertilized channel catfish eggs were incubated at five constant water temperatures: 4, 11. 16. 21, and 26 C. Low-temperature incubation of catfish eggs extended the embryonic period at 16 (244%) and 21 C (56%) when compared to the control hatchery incubation temperature of 26 C. All eggs incubated at 4 and 11 C died within 24–48 h. Developmental stage had a significant ( P > 0.05) effect on percent hatch at 16, 21, and 26 C. Eggs held at 16 C prior to embryonic axis formation died within 48 h. Larvae from eggs hatched at 16 C were incompletely developed and died upon acclimation to 26 C for growth tests. Growth of fry reared at 26 C, following egg incubation at 21 C. paralleled that of fry from eggs incubated at 26 C. The underdevelopment of fry at 16 C combined with the significant effect of egg stage on survival at this temperature suggests that 16 C is below the lower thermal tolerance limit for normal development in this species. The period prior to the formation of the embryonic axis may be considered a vulnerable stage in channel catfish development. Increasing the embryonic period through low temperature incubation would increase the duration of juvenile availability for researchers and commercial operations.     

Extenders and cryoprotectants for cooling and freezing of piracanjuba (Brycon orbignyanus) semen, an endangered Brazilian teleost fish

The aim of this study was to develop a protocol for semen storage of piracanjuba (Brycon orbignyanus) by both cool storage at 4 °C and cryopreservation at − 196 °C. Semen was diluted in some fish semen extenders (Exp. 1) or in extenders combined with the antibiotic gentamycin sulfate (Exp. 2) and stored at 4 °C. Sperm motility was estimated every 24 h. Then, the effects of egg yolk (0 and 5%), cryoprotectants (dimethyl sulphoxide — DMSO, methanol, and methylglycol) and extenders (NaCl 154 mM, BTS™ Minitub and M III™ Minitub) on semen cryopreservation were evaluated (Exp. 3). Semen was added to each of eighteen cryosolutions (2 yolk concentrations × 3 cryoprotectants × 3 extenders), aspirated into 0.5-mL straws, frozen in nitrogen vapor (Taylor-Wharton, CP 300, “dry shipper”) and stored at − 196 °C. Sperm motility was evaluated after thawing at 60 °C-water bath for 8 s. The three cryosolutions that produced the highest post-thaw sperm motility were used again to freeze semen. Post-thaw semen quality was then evaluated under three tests: sperm motility, the percentage of live spermatozoa and hatching rate (Exp. 4). Piracanjuba semen diluted (1:10 total volume) in NaCl 200 mM or in Saad solution (NaCl 200 mM, Tris 30 mM) maintained motility above 35% for as long as 7 days, at 4 °C. Motility of only 7% was observed on undiluted semen after 3 days at 4 °C. There was neither beneficial nor detrimental effect of gentamycin on sperm motility at 250 μg/mL. Egg yolk addition to the cryosolution was beneficial in samples cryopreserved in NaCl 154 mM and in M III™, but detrimental for samples cryopreserved in BTS™. Methylglycol was the most effective cryoprotector compared to DMSO and methanol. Motility and percentage of live spermatozoa were similar among semen cryopreserved in NaCl–yolk, M III™–yolk and BTS™, all containing 10% methylglycol, but lower than fresh control. Hatching rates of eggs fertilized with sperm cryopreserved in NaCl–yolk or BTS™ were higher than for eggs fertilized with sperm cryopreserved in M III™–yolk, but lower than control fertilizations. The semen cryopreservation protocols developed here will be used to set up a gene bank for endangered piracanjuba populations.     

Commensal bacteria and their possible relationship to the mortahty of incubating salmonid eggs

Abstract. Eggs of female rainbow trout, Oncorhynchus mykiss (Walbaum), were fertilized and subsequently exposed to high eoncentrations of Pseudomonas fluorescens or Cytophaga sp. (10⁶ cells/ml) for 45 min during water hardening. Eggs were placed in horizontal hatchery trays alongside sterile water-treated eggs and mortality rates were observed daily throughout incubation. Numbers and species of bacteria present on egg surfaces and within egg contents were monitored at the green stage (unfertilized), after hardening, and at 12h, 7 days, 14 days, 21 days and 28 days incubation. Surfaces of the incubating eggs of all groups, irrespective of treatment at water hardening, were eventually colonized by considerable numbers of Cytophaga sp., P. fluorescens and to a lesser extent Pseudomonas sp. and Aeromonas hydrophila. Few bacteria were recovered from inside eggs. Overall, no correlation was found between bacteria on the egg surface and egg death. However, a significantly higher 'rate' of egg death during early stages of incubation was found amongst eggs exposed to P. fluorescens at water hardening. Thus, high numbers of P. fluorescens on egg surfaces during initial stages of incubation might pose a potential threat to egg survival. Therefore, it is suggested that strenuous efforts are made at time of hardening to obtain the purest available source of water.     

Effect of low water temperature on viral replication of white spot syndrome virus in Procambarus clarkii

This study evaluated the effect of low water temperature (10 ± 1 °C) on viral infection and replication of white spot syndrome virus (WSSV) in crayfish, Procambarus clarkii, under standardized conditions. Crayfish were (i) maintained at 24 ± 1 °C before challenge and 10 ± 1 °C afterwards, or (ii) maintained at 10 ± 1 °C before challenge and 24 ± 1 °C afterwards. No mortality was observed when crayfish were held at 10 ± 1 °C after challenge, but mortality reached 100% when they were transferred to 24 ± 1 °C. Competitive PCR showed that viral levels at 10 ± 1 °C rose from 10⁶ to 10⁸ copies/mg of gill tissues, while at 24 ± 1 °C levels increased from 10⁶ to 10¹⁰ copies/mg of gill tissues during the same time interval. These results showed that a low water temperature of 10 ± 1 °C could reduce viral replication when compared to 24 ± 1 °C but could not prevent it.     

Some effects of temperature and salinity on laboratory-reared eggs and larvae of Polydactylus sexfilis (Pisces: Polynemidae)

Effects of temperature and salinity on eggs and yolksac larvae of Polydactylus sexfilis (Cuvier and Valenciennes) were examined in laboratory experiments. Data on developmental rates as influenced by temperature are presented. Larval length at 95% yolksac absorption was maximized between 23.8 and 28.6°C. Based on the development of functional eyes and jaws, larvae were judged capable of feeding before the yolk was completely absorbed. Larvae incubated at intermediate temperatures also had larger amounts of yolk remaining when eyes and jaws were judged functional. Temperature and salinity effects on hatching success, survival at the end of the yolksac stage, and morphological abnormalities were studied in a 10 × 5 (temperature × salinity) array of treatments. In 34‰ sea water, normalized larval survival at the end of the yolksac stage was greater than 50% between temperatures of 21.9 and 28.0°C. Larval survival decreased at lower temperatures and salinities. Proportions of abnormal larvae increased at temperature and salinity extremes, and normal development was maximized between 26 and 34‰. Larvae (74 h after fertilization) were more tolerant to extreme high temperatures than were newly fertilized eggs. Upper salinity tolerance limits of 42-h larvae were greater at 26.2°C than at 23.5 or 29.2°C, and lower salinity was less tolerated at the two extreme temperatures. Based on the results, recommended temperatures and salinities for rearing P. sexfilis eggs and early larvae are 24–28°C and 26–34‰.     

First multi-generation culture of the tropical cuttlefish Sepia pharaonis Ehrenberg, 1831

Sepiapharaonis, the pharaoh cuttlefish was cultured through multiplegenerations in the laboratory (5 consecutive generations) using closed,recirculating water filtration systems. The eggs of the original parentalgeneration (G_P) were spawned by a wild caught Gulf of Thailandfemale in alocal fisheries laboratory, then packed and shipped air cargo to Texas wherehatching occurred. The culture temperature ranged 25°–28°C, except for one generation that was chilled intentionallyto21 °C and then warmed to 25 °C after 9.6months. Spawning occurred as early as day 161. Spawning output was high in allgenerations except the group that was cultured at 21 °C. Eggfertility was low in captivity (< 20%), but hatchling survival was high(>70%). The average egg incubation time was 13.6 d at 25–28°C. The largest spawn resulted in 600 viable hatchlings andthesmallest resulted in 11 hatchlings. The cuttlefish ate a wide variety ofestuarine crustaceans and fishes as well as frozen shrimp. There were noapparent disease problems since survival from hatching to maturity was over70%.The average life span for cuttlefish cultured at 25–28°Cwas 8.9 months and 12.3 months at 21 °C. Size at hatching wasmeasured for fourth generation (G₄) hatchlings; the mean weight athatching was 0.103 g and the mean mantle length was 6.4mm. The largest cuttlefish cultured was a male 300 mmML and 3,045 g; the oldest cuttlefish lived 340 d.This cuttlefish species presents an excellent choice for commercial mariculturebecause of its rapid growth, short life span, tolerance to crowding andhandling, resistance to disease and feeding habits.     

The effect of temperature and duration of incubation on the hatching of diapause eggs of Centropages hamatus (Lilljeborg) (Copepoda, Calanoida)

Diapause eggs of Centropages hamatus were used to investigate the effect of temperature and duration of incubation on egg hatching. Eggs were incubated for 10, 12, 14, 16, 20, 24, 28, 32, 36 and 40 h at 15°C and 14L–10D. After incubation for the designated period, eggs were transferred to 25°C and monitored periodically to determine egg hatching. Control eggs were incubated solely at 15°C and monitored for egg hatching. The greatest daily hatching success of eggs occurred within 1 or 2 days after transfer from 15°C to 25°C, while the controls required 3–4 days. The cumulative hatching success of eggs was significantly lower than the control, with the exception of eggs held for at least 36 h at 15°C before transfer to 25°C. These results indicate that overall time to hatching of diapause eggs of C. hamatus can be reduced by transferring the eggs to a higher temperature, for example, 25°C, following a minimum period of time (36 h) at reduced temperature, for example, 15°C. Exposure to 15°C for only 10 h does not appear to be sufficient to result in any subsequent hatching at higher temperature.     

Effects of chloramphenicol, erythromycin, and furazolidone on growth of Isochrysis galbana and Chaetoceros gracilis

This study focused on determining the effects of antibiotics on microalgae used as food for scallop larvae. Six different dose levels of chloramphenicol, erythromycin, and furazolidone were added to cultures of Isochrysis galbana and Chaetoceros gracilis. An in vivo experiment was subsequently conducted to determine the effect of chloramphenicol and erythromycin on larval survival of the Pacific calico scallop Argopecten ventricosus in tanks and on the population of its associated bacteria. Results showed that growth of I. galbana was not significantly affected by chloramphenicol or erythromycin at the test doses of 0.5, 1.0, 3.0, 6.0, 9.0, and 12.0 mg/l. C. gracilis was significantly sensitive to erythromycin and chloramphenicol at doses higher than 0.5 and 3.0 mg/l, respectively. Furazolidone inhibited the growth of both I. galbana and C. gracilis at all test doses. Results showed that exposure of scallop larvae to a dose of 6 mg/l chloramphenicol or erythromycin did not significantly affect growth of I. galbana, significantly enhanced survival of the scallop larvae, and inhibited the growth of Vibrio spp. in tanks. This study demonstrated the adverse effect of chloramphenicol, erythromycin and furazolidone on I. galbana and C. gracilis microalgae but the positive effect on survival of the scallop larvae, decreasing associated bacterial population.     

Spawning, early development and first feeding in the gobiid fish Priolepis nocturna

The reproductive behavior, embryonic development and early larvae of Priolepis nocturna are described. Three pairs of P. nocturna began spawning 41 days after acquisition and maintained a 5–10 day spawning cycle lasting beyond several months. Spawning was initiated by the female who signaled her readiness to spawn by displaying to the male. Egg clutch size averaged 1578 ± 51.23 eggs and ranged from 268 to 3121. Egg length averaged 0.82 ± 0.01 mm total length (TL) and ranged from 0.75 to 0.90 mm. Egg width averaged 0.51 ± 0.51 mm total width (TW) and ranged from 0.49 to 0.52 mm. Fertilized eggs were ovoid in shape and attached to the ceiling of provided shelters via adhesive filaments at the proximal end. Hatching rates averaged 97.3 ± 0.51% and ranged from 91.9 to 99.8%. Larvae measuring 1.89 ± 0.04 mm TL hatched 121 ± 0.5 h post fertilization and did not rotate position prior to hatching. Skeletal elements of the chondrocranium were simplistic and dominated by the hyoid, hyomandibulosymplectic cartilage, ethmoid and Meckel's cartilage in first feeding larvae. No elements were added to the cranial architecture by 5 days post hatch (DPH) when larvae measured 2.05 ± 0.04 mm TL. First feeding larvae consumed only dinoflagellates and tintinnids suggesting that feeding was constrained by a poorly developed feeding mechanism. Embryology and larval development are described to 5 DPH.     

Induction of triploidy in the turbot (Scophthalmus maximus): II. Effects of cold shock timing and induction of triploidy in a large volume of eggs

Triploidy was induced in the turbot (Scophthalmus maximus, L.) by applying cold shocks shortly after fertilization. The combined effects of the timing of cold shock commencement after fertilization, cold shock duration and cold shock temperature were investigated. Ploidy was assessed by counting the number of nucleoli per nucleus (NOR) in larvae and also by measuring erythrocyte size in juveniles. A clear peak in triploidy induction was obtained when shocks were started between 6 and 7 min after fertilization at a pre-shock temperature of 13–14°C. With this timing, shocks of 20-min duration at 0°C gave >90% triploidy, with survival about 80% of the untreated controls. In order to ensure both high triploidy rates and high survival, it was necessary to carefully maintain the water temperature just below 0°C. Experiments with small and large volumes of eggs were performed in order to determine how changes in the relative volumes of eggs and chilled water could affect survival and triploidy induction. The best combination to induce triploidy in the turbot was as follows: shock commencement 6.5 min after fertilization, shock duration 25 min, and shock temperature between 0 and −1°C. With this combination, 100% triploidy could consistently be induced with survival 60% of the untreated control. This was successfully applied to a large volume of eggs (300 ml; 1 ml 800 eggs) in order to mass-produce triploid turbot. Triploids had lower survival rate than diploids at hatching but similar thereafter, with the ability to complete the different stages of larval rearing, indicating the viability to produce triploid turbot under farming conditions.     

First multi-generation culture of the tropical cuttlefish Sepia pharaonis Ehrenberg, 1831

Sepiapharaonis, the pharaoh cuttlefish was cultured through multiplegenerations in the laboratory (5 consecutive generations) using closed,recirculating water filtration systems. The eggs of the original parentalgeneration (G_P) were spawned by a wild caught Gulf of Thailandfemale in alocal fisheries laboratory, then packed and shipped air cargo to Texas wherehatching occurred. The culture temperature ranged 25°–28°C, except for one generation that was chilled intentionallyto21 °C and then warmed to 25 °C after 9.6months. Spawning occurred as early as day 161. Spawning output was high in allgenerations except the group that was cultured at 21 °C. Eggfertility was low in captivity (< 20%), but hatchling survival was high(>70%). The average egg incubation time was 13.6 d at 25–28°C. The largest spawn resulted in 600 viable hatchlings andthesmallest resulted in 11 hatchlings. The cuttlefish ate a wide variety ofestuarine crustaceans and fishes as well as frozen shrimp. There were noapparent disease problems since survival from hatching to maturity was over70%.The average life span for cuttlefish cultured at 25–28°Cwas 8.9 months and 12.3 months at 21 °C. Size at hatching wasmeasured for fourth generation (G₄) hatchlings; the mean weight athatching was 0.103 g and the mean mantle length was 6.4mm. The largest cuttlefish cultured was a male 300 mmML and 3,045 g; the oldest cuttlefish lived 340 d.This cuttlefish species presents an excellent choice for commercial mariculturebecause of its rapid growth, short life span, tolerance to crowding andhandling, resistance to disease and feeding habits.