The effect of temperature on the resting and post-exercise metabolic rates and aerobic metabolic scope in shortnose sturgeon <Emphasis Type="Italic">Acipenser brevirostrum</Emphasis>

The effects of acclimation temperature (15, 20, 25 °C) on routine oxygen consumption and post-exercise maximal oxygen consumption rates (MO₂) were measured in juvenile shortnose sturgeon (Acipenser brevirostrum LeSueur, 1818). The routine MO₂ of shortnose sturgeon increased significantly from 126.75 mg O₂ h^?1 kg^?1 at 15 °C to 253.13 mg O₂ h^?1 kg^?1 at 25 °C. The temperature coefficient (Q ₁₀) values of the routine metabolic rates ranged between 1.61 and 2.46, with the largest Q ₁₀ values occurring between 15 and 20 °C. The average post-exercise MO₂ of all temperature groups increased to a peak value immediately following the exercise, with levels increasing about 2-fold among all temperature groups. The Q ₁₀ values for post-exercise MO₂ ranged from 1.21 to 2.12, with the highest difference occurring between 15 and 20 °C. Post-exercise MO₂ values of shortnose sturgeon in different temperature groups all decreased exponentially and statistically returned to pre-exercise (resting) levels by 30 min at 15 and 20 °C and by 60 min at 25 °C. The aerobic metabolic scope (post-exercise maximal MO₂-routine MO₂) increased to a maximum value ～156 mg O₂ h^?1 kg^?1 at intermediate experimental temperatures (i.e., 20 °C) and then decreased as the temperature increased to 25 °C. However, this trend was not significant. The results suggest that juvenile shortnose sturgeon show flexibility in their ability to adapt to various temperature environments and in their responses to exhaustive exercise.     

Osmoregulation in juvenile Chinese sturgeon (<Emphasis Type="Italic">Acipenser sinensis</Emphasis> Gray) during brackish water adaptation

The osmoregulation capabilities of 7-month-old juvenile Chinese sturgeon (Acipenser sinensis Gray) (128.8 ± 15 g) transferred directly from fresh water (0‰, 46 mOsmol kg⁻¹) to brackish water (10‰, 273 mOsmol kg⁻¹) were studied over a 20-day period. Changes in serum osmolarity, chloride (Cl⁻), sodium (Na⁺), potassium (K⁺) and calcium (Ca²⁺) ion concentrations, as well as gill and spiral valve Na⁺,K⁺-ATPase activities were measured at 3, 12, 24, 72, 216 and 480 h after transfer to BW. The serum osmolarity and ion concentrations (Na⁺, Cl⁻ and Ca²⁺) increased immediately after the transference to BW, reaching maximum at 24 h and returned to a new steady state at 216 h, while the FW control group maintained basal levels which showed lower (P < 0.05) than the BW group. Gill Na⁺,K⁺-ATPase activity of BW group exhibited an abrupt decrease in the first 3 h after transfer, but began to increase at 3 h, reaching a peak value at 24 h, and returned to a new steady state at 216 h. The differences between gill Na⁺,K⁺-ATPase activity of BW and FW fish were significant (P < 0.05) after 12 h. In contrast, Na⁺,K⁺-ATPase activity of the spiral valve showed transient increase after transference from FW to BW, and then decreased rapidly at 3 h, reaching the lowest at 24 h after transference. At 216 h after exposure to BW, Na⁺,K⁺-ATPase activities of the spiral valve increased slowly to the levels of FW control. The results of our study indicate the existence of hyposmoregulatory adaptive mechanisms in 7-month-old juvenile Chinese sturgeon which enable this fish to acclimate itself successfully to brackish water.     

Salinity effects on osmoregulation and gill morphology in juvenile Persian sturgeon (<Emphasis Type="Italic">Acipenser persicus</Emphasis>)

The effect of abrupt and 5-day gradual salinity transfers from freshwater (FW) to 11 ‰ Caspian Sea brackish water (BW) was investigated in juvenile Persian sturgeon Acipenser persicus with three different weight groups: 1–2 g (1.62 ± 0.27 g), 2–3 g (2.55 ± 0.41 g) and 3–5 g (4.28 ± 0.76 g). Mortality rates, blood osmotic pressure, gill morphology and branchial Na⁺, K⁺-ATPase (NKA) activity were measured 4 and 10 days after abrupt transfer and 9 and 15 days after the initial gradual transfer (i.e. 4 and 10 days after reaching Caspian Sea salinity). Fish under 3 g could not survive increased salinity, and the blood osmotic pressure of the remaining surviving fish increased and remained elevated. However, heavier fish were able to survive and successfully acclimate, even to rapid salinity change with osmotic pressure reduced to Caspian Sea osmolality levels. At the gill level, the developmental increase in chloride cell volume and a higher NKA content most probably allow juveniles weighing more than 2 g to sharply increase NKA activity if the fish are transferred to BW. The rapid chloride cell proliferation occurring with increased salinity should strengthen this acclimation response. Therefore, a drastic physiological change occurs when fish weigh more than 2 g that allows migration to higher salinities. The triggering signal on chloride cells must be further investigated in order to optimize this functional step.     

Farming evaluation of the brownbow triploid hybrid (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis><Emphasis Type="Italic">Salmo trutta</Emphasis>)

Triploid hybrids between female rainbow trout (Oncorhynchus mykiss Walbaum) and male brown trout (Salmo trutta L.) were tested for farming performances, with reference to parental species. The main drawback of hybrids lay in embryonic and larval mortalities, amounting to 60% on average, and displaying a large variability between spawns. Further survival was inferior to that of diploid, but similar to that of triploid rainbow trout. Hybrid body weight was intermediate between weights of rainbow and brown trout of the same age, mainly as a consequence of differences in precocious growth. Analysis of relative growth rates from 6 to 18 months showed that hybrids were surpassed by rainbow controls in common rearing, but not in separate rearing. Hybrid behaviour was similar to that of rainbow trout. These results are discussed in the scope of providing fisheries managers with original and sterile game fishes. An erratum to this article can be found at .     

Hematology of great sturgeon (<Emphasis Type="Italic">Huso huso</Emphasis> Linnaeus, 1758) juvenile exposed to brackish water environment

The effect of environmental salinity on hematological parameters of great sturgeon Huso huso juveniles was studied. Five-month-old juveniles (mean body weight 28.3 ± 2.1 g) were subjected to 0, 3, 6, 9, and 12 ppt salinities. The hematological parameters were assessed after a period of 20 days rearing at these salinities. After transfer from fresh water to brackish water, red blood cells, hematocrit, haemoglobin and mean corpuscular haemoglobin decreased, but mean corpuscular volume increased. Mean corpuscular haemoglobin concentration, white blood cells, monocyte counts, and eosinophil counts showed no significant variations with increase in environmental salinity. An increase was found in lymphocyte counts according to the increase of salinity from 0 to 12 ppt, while the fresh water control group maintained basal levels. Decrease in neutrophil counts was observed in great sturgeon with increase in environmental salinity. These data show significant effect of salinity on the blood parameters of great sturgeon.     

Tissue-specific daily variation in the oxidative status of sturgeon (<Emphasis Type="Italic">Acipenser naccarii</Emphasis>) and rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>): a comparative study

The oxidative status is associated with animal lifespan, metabolism, activity and circadian rhythms. The objective of this work is to study the time course of the oxidative status over a daily cycle in the plasma, liver and brain, and the changes in the plasma cortisol levels of sturgeon and trout. The knowledge of daily oxidative status will provide a better understanding of the trout and sturgeon physiology and adequate maintenance and food supply of farmed fish in relation to photoperiod. Superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione transferase, DT-diaphorase activities, lipid peroxidation and cortisol were measured. Our results showed that the antioxidative enzyme activities and lipid peroxidation in the liver of trout and sturgeon changed through the day, with increased levels in lipid peroxidation of liver in the dark period for sturgeon. This could be related to the different activity time in both species, an issue to be taken into account when designing the guidelines of the maintenance of these species in fish farms. On the contrary, there was not clear influence of the daily rhythms on brain oxidative status. The higher efficiency of the antioxidant defences in the brain of sturgeon, which displays less lipid peroxidation and higher antioxidative activity, could be related to its longer life expectancy. The absence of any apparent daily rhythm in the plasma cortisol levels in sturgeon could indicate a lower susceptibility to stress, and that mechanisms involved in cortisol secretion in chondrosteans could be different from that described for teleosteans.     

Effects of temperature and fatigue on the metabolism and swimming capacity of juvenile Chinese sturgeon (<Emphasis Type="Italic">Acipenser sinensis</Emphasis>)

Chinese sturgeon (Acipenser sinensis) is a critically endangered species. A flume-type respirometer, with video, was used to conduct two consecutive stepped velocity tests at 10, 15, 20, and 25 °C. Extent of recovery was measured after the 60-min recovery period between trials, and the recovery ratio for critical swimming speed (U _crit) averaged 91.88% across temperatures. Temperature (T) effects were determined by comparing U _crit, oxygen consumption rate (MO ₂), and tail beat frequency (TBF) for each temperature. Results from the two trials were compared to determine the effect of exercise. The U _crit occurring at 15 °C in both trials was significantly higher than that at 10 and 25 °C (p < 0.05). The U _crit was plotted as a function of T and curve-fitting allowed calculation of the optimal swimming temperature 3.28 BL/s at 15.96 °C (trial 1) and 2.98 BL/s at 15.85 °C (trial 2). In trial 1, MO ₂ increased rapidly with U, but then declined sharply as swimming speed approached U _crit. In trial 2, MO ₂ increased more slowly, but continuously, to U _crit. TBF was directly proportional to U and the slope (dTBF/dU) for trial 2 was significantly lower than that for trial 1. The inverse slope (tail beats per body length, TB/BL) is a measure of swimming efficiency and the significant difference in slopes implies that the exercise training provided by trial 1 led to a significant increase in swimming efficiency in trial 2.     

Effects of dietary <Emphasis Type="Italic">myo</Emphasis>-inositol on growth,chemical composition and plasma chemistry of Amur sturgeon <Emphasis Type="Italic">Acipenser schrenckii</Emphasis>

The present study was conducted to demonstrate the dietary myo-inositol requirement and its effects on the growth, proximate composition and blood chemistry of Amur sturgeon (Acipenser schrenckii). Triplicate groups of 30 fish (initial weight 11.90?±?0.12 g) were fed different diets containing graded levels of myo-inositol (28.75, 127.83, 343.83, 565.81, 738.15 and 936.28 mg kg^?1) until satiation for 56 days. The fish were weighed after a 24-h fast, and six fish were used for whole body composition analysis. Further, the liver and muscle were sampled from another six fish for lipid analysis. The blood and liver were sampled from the remaining six fish for haematology and fatty acid analysis. The weight gain of fish increased with myo-inositol content, from the 28.75- to 343.83-mg kg^?1 myo-inositol treatment groups, and then stabilised. The liver lipid content and hepatosomatic index decreased significantly from 21.91 to 19.14% and from 3.20 to 2.76% with increased dietary myo-inositol supplementation, respectively. The whole body lipid content generally decreased from 6.33 to 5.55%. The content of liver-saturated fatty acids decreased significantly (28.13%) in the 936.28-mg kg^?1 treatment group. The content of plasma non-esterified fatty acids increased with the increase in dietary myo-inositol supplementation from 0.77 to 1.17 mmol L^?1, whereas the content of triglycerides significantly decreased from 4.62 to 3.28 mmol L^?1. In conclusion, the optimum myo-inositol requirement was found to be 336.1 mg kg^?1, based on weight gain in a two-slope quadratic broken-line model.     

Anesthesia induces stress in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>), Atlantic cod (<Emphasis Type="Italic">Gadus morhua</Emphasis>) and Atlantic halibut (<Emphasis Type="Italic">Hippoglossus hippoglossus</Emphasis>)

Stress in response to anesthesia with benzocaine, MS-222, metomidate and isoeugenol was studied in Atlantic salmon (Salmo salar), Atlantic halibut (Hippoglossus hippoglossus), and Atlantic cod (Gadus morhua) with no concomitant stress from handling or confinement in association with anesthesia or sampling. All of the anesthetics tested induced a stress response in all species, displayed by a release of cortisol to the water. MS-222 anesthesia elicited the highest cortisol release rates, reaching maximum levels 0.5 h post-exposure and returning to basal levels after 3–4 h. Benzocaine anesthesia caused a bimodal response where the initial peak in cortisol release rate was followed by a second increase lasting towards the end of the trial (6 h). This bimodality was more profound in Atlantic salmon than in Atlantic halibut and Atlantic cod. Metomidate anesthesia induced the lowest release of cortisol of the agents tested in both Atlantic halibut and Atlantic cod, but resulted in a bimodal response in Atlantic salmon where the initial increase in cortisol release was followed by a larger increase peaking at 2–2.5 h post exposure before returning to basal levels after 5 h. The stress induced in Atlantic salmon by isoeugenol anesthesia resembled that of MS-222, but did not reach the same elevated level. Overall, the cortisol release was most profound in Atlantic salmon followed by Atlantic halibut and Atlantic cod.     

Effect of dietary macronutrient proportion on intermediate metabolism and oxidative status in sturgeon (<Emphasis Type="Italic">Acipenser naccarii</Emphasis>) and trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>): comparative study

Three isoenergetic diets varying the proportion of dietary energy supplied by each of the macronutrients (carbohydrate, lipid, or protein) were delivered, to farmed sturgeon (Acipenser naccarii) and trout (Oncorhynchus mykiss), to test the possible effects on the intermediate metabolism and oxidative status in liver, white muscle, and heart. In trout, there is an adaptive metabolic response to an increase in lipids and carbohydrates in the diet. However, this does not happen in the sturgeon. These differences may be due to different dietary habits of both species. In terms of oxidative status, only the liver displayed oxidative stress in both species, showing an increase in the lipid peroxidation and antioxidant enzyme activities after feeding with the high-lipid and high-protein diet.     

<Emphasis Type="Italic">Neoheterobothrium hirame</Emphasis> (Monogenea) alters the feeding behavior of juvenile olive flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

We monitored feeding behavior and survival of starved juvenile olive flounder experimentally infected with the gill monogenean Neoheterobothrium hirame. Infected flounder increased amount of the time spent in the water column by 117% when trying to capture live mysids, Neomysis sp. They also showed different feeding patterns from those of uninfected fish and made fewer attacks towards prey during one feeding attempt. Although the average numbers of mysids captured by individuals were similar between infected and uninfected fish, heavily infected fish tended to catch less prey. These results indicate that N. hirame reduces the feeding efficiency of the host for capturing live prey and possibly makes them more vulnerable to predation during feeding. We could not detect any obvious difference in survival rates between uninfected, lightly and heavily infected fish during 3 months of starvation. There was no evidence that starvation makes fish more susceptible to N. hirame. The present study provides first experimental evidence that N. hirame affects feeding behavior of juvenile olive flounder and supports the idea that this parasite indirectly reducing the host’s survival and may be responsible for the recent reduction of the flounder population in Japan.     

Recent expansion of Northeast Atlantic and Mediterranean populations of <Emphasis Type="Italic">Melicertus</Emphasis> (<Emphasis Type="Italic">Penaeus</Emphasis>) <Emphasis Type="Italic">kerathurus</Emphasis> (Crustacea: Decapoda)

We analysed the genetic diversity of Melicertus kerathurus (Penaeidae), a commercially valuable penaeid shrimp that is distributed in the Mediterranean Sea and eastern Atlantic Ocean. We examined the polymorphism of a 494 bp DNA segment of the mitochondrial COI region in 173 individuals, sampled in nine Mediterranean and two Atlantic samples, covering the whole range of the species from the tropical waters of the Gulf of Guinea to the eastern part of the Mediterranean Sea. The mean nucleotide and haplotype diversities were π = 0.00275 and h = 0.718, respectively, for the global data set, with the highest values occurring in the African samples and the lowest in the Adriatic Sea. A clear sample differentiation was found (F _st = 0.194), but this did not reflect a geographical pattern and there were only faint traces of an Atlantic–Mediterranean subdivision. Mismatch analysis and a high significant negative value of Tajima’s D suggested that M. kerathurus is not at mutation drift-equilibrium, but underwent a recent expansion after a period of low effective sample size. A postglacial recolonisation of the Mediterranean from an Atlantic refuge could be hypothesised based on these data.     

Monitoring of <Emphasis Type="Italic">Francisella noatunensis</Emphasis> subsp. <Emphasis Type="Italic">orientalis</Emphasis> in farmed Nile tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>) in Brazil

Francisella noatunensis orientalis is a bacterium that causes emerging bacteriosis in Nile tilapia (Oreochromis niloticus) in many parts of the world, including Brazil. It is a non-motile, Gram-negative, strictly aerobic, facultative intracellular coccobacillus. This species of bacteria is responsible for low to high mortality in fish farms, causing economic losses for fish farmers. This study aimed to detect the presence of F. noatunensis orientalis using qPCR (real-time polymerase chain reaction) and to describe lesions caused by the bacterium in O. niloticus in Brazilian aquaculture. For this purpose, 360 fish from six fish farms (30 per farm) were sampled at two time points (n = 180 per sampling). Necropsies and histopathology were performed for lesion observation, in addition to qPCR and sequencing for detection and identification of Francisella species. Environmental data were collected using a multiparameter sonde YSI EXO2. All measured limnological variables were within the optimum range for cultivation of Nile tilapia. The major lesions present were melanization of the skin, splenomegaly, granulomas, and inflammatory cell responses. The prevalence of francisellosis varied from 0 to 86.66% between time periods and fish farms analyzed, and an outbreak was observed during the second sampling period. This study describes the prevalence of francisellosis in O. niloticus and reports that the lesions found are not exclusively associated with this bacterial disease.     

Potential of cinnamon (<Emphasis Type="Italic">Cinnamomum verum</Emphasis>) oil to control <Emphasis Type="Italic">Streptococcus iniae</Emphasis> infection in tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>)

In this study, four essential oils—cinnamon oil, leech lime oil, lemongrass oil, and turmeric oil—were examined for their antimicrobial activities against Streptococcus iniae, a bacterium that is pathogenic in fish, in which it causes streptococcosis. Cinnamon oil was the most potent antimicrobial agent among these oils, with a minimal inhibitory concentration (MIC) of 40 μg/ml. By using gas chromatography–mass spectrometry (GC–MS), it was found that the major components of cinnamon oil were cinnamaldehyde (90.24), limonene (2.42%), cinnamyl acetate (2.03%), linalool (1.16%), and α-terpineol (0.87%). Of these compounds, only cinnamaldehyde exhibited antimicrobial activity against S. iniae, with an MIC of 20 μg/ml. In an in vivo trial, no mortality was apparent in fish fed on fish diets supplemented with 0.4% (w/w) of cinnamon oil and with 0.1% (w/w) of oxytetracycline 5 days prior to infection with S. iniae. These results indicate that cinnamon oil had a protective effect on experimental S. iniae infection in tilapia, and thus has the potential to replace the antibiotics used to control this disease.     

Growth and survival of grouper <Emphasis Type="Italic">Epinephelus</Emphasis><Emphasis Type="Italic">coioides</Emphasis> (Hamilton) larvae fed free-living nematode <Emphasis Type="Italic">Panagrellus redivivus</Emphasis> at first feeding

The free-living nematode, Panagrellus redivivus, was tested as live food for grouper Epinephelus coioides larvae during the first feeding stage. A series of experiments were conducted to determine the acceptability of the free-living nematodes in grouper larvae at first feeding, the optimum nematode density and the response of the larvae to nutritionally enriched nematode. All experiments were conducted in 200-L conical tanks filled with 150-L filtered seawater and stocked at 15 larvae L⁻¹. Duration of feeding experiments was up to day 21 (experiment 1) and 14 days (experiment 2 and 3). Brachionus plicatilis and Artemia (experiment 1) and Brachionus plicatilis alone (experiment 2 & 3) was used as the control treatment. Observations indicated that the grouper larvae readily fed on free-living nematodes as early as 3 days posthatching, the start of exogenous feeding. Optimum feeding density for the larvae was 75 nematodes ml⁻¹. The enrichment of cod liver oil or sunflower oil influenced the total lipids and n-3 highly unsaturated fatty acids of P. redivivus, which in turn influenced those of the grouper larvae, however, growth and survival of the larvae were not affected (P > 0.05). The results from this investigation showed that the nematode, P. redivivus, can be used as first live food for grouper larvae from the onset of exogenous feeding until they could feed on Artemia nauplii.     

<Emphasis Type="Italic">Astragalus membranaceus</Emphasis> (AM) enhances growth performance and antioxidant stress profiles in bluegill sunfish (<Emphasis Type="Italic">Lepomis macrochirus</Emphasis>)

This study was designed to assess the potential effects of Astragalus membranaceus (AM) on the growth performance and antioxidative stress response in bluegill sunfish (Lepomis macrochirus) exposed to challenging cold water temperature conditions. In this regard, fish with an average weight of 43 ± 1 g were divided into four groups and fed daily with an AM-free diet (control), and 1.5, 3, and 4.5 % (w/w) AM-incorporated diets for an 8-week period. Oxidative stress response, biochemical, and growth parameters were measured, and subgroups of fish were exposed to the outside challenging cold pond water temperature (1.6–9.9 °C) with an average of 7.0 ± 0.1 °C beyond the optimal temperature. The results showed that incorporating AM in the diet significantly improved growth performance parameters (body mass gain, specific growth rate, length, condition factor, and feed conversion ratio) and biochemicals (aspartate aminotransferase and alanine transaminase activities, and glucose and cortisol concentrations). In addition, markedly up-regulated superoxide dismutase, glutathione peroxidase, and catalase activities were observed in AM-treated fish groups over the control. Conclusively, feeding AM diets significantly increased (P < 0.05) growth performance and antioxidative stress profiles throughout the entire experiment, and this increase was much more pronounced at 8 weeks after the water temperature began to rise, which can be related to the nature of Bluegill fish as it is known to be a warm water fish. These findings are considered to be of great importance for sustainable aquaculture.     

Spontaneous polyploidization in critically endangered <Emphasis Type="Italic">Acipenser mikadoi</Emphasis>

We investigated the source of spontaneous polyploidization in the critically endangered Acipenser mikadoi. Fourteen sib progeny of A. mikadoi and 11 hybrids between an A. mikadoi female and a Huso dauricus male, all showing atypically high ploidy, were analysed. Parent assignment based on five highly polymorphic microsatellite markers confirmed spontaneous duplication of maternal chromosome sets via retention of the second polar body to be the source of spontaneous polyploidization. To our knowledge, this provides the first evidence of the maternal origin of spontaneous polyploidization in A. mikadoi. Factorial correspondence analysis of the multilocus microsatellite genotypes placed the parent fish of spontaneous polyploids in clearly delineated clusters of A. mikadoi and H. dauricus, and parent fish had mitochondrial control region haplotypes corresponding to their presumed species. Thus, parent fish were confirmed to be of pure genetic origin, and hybridization did not promote the observed spontaneous polyploidization.     

Spatial learning-induced <Emphasis Type="Italic">egr</Emphasis>-<Emphasis Type="Italic">1</Emphasis> expression in telencephalon of gold fish <Emphasis Type="Italic">Carassius auratus</Emphasis>

The immediate-early gene (egr-1) expression was used to examine the neuron’s response in telencephalon of goldfish during spatial learning in small space. Fishes were pre-exposed in the experimental apparatus and trained to pick food from the tray in a rectangular-shaped arena. The apparatus was divided into identical compartments comprising three gates to provide different spatial tasks. After the fish learned to pass through the gate one, two more gates were introduced one by one. Fish made more number of attempts and took longer time (P < 0.05) to pass through the first gate than the gate two or three. This active learning induces the expression of egr-1 in telencephalon as established by western blot analysis. Subsequently, the fish learn quickly to cross the similar type of second and third gate and make fewer errors with a corresponding decline in the level of egr-1 expression. As the fish learned to pass through all the three gates, third gate was replaced by modified gate three. Interestingly, the level of egr-1 expression increased again, when the fish exhibit a high exploratory behavior to cross the modified gate three. The present study shows that egr-1 expression is induced in the telencephalon of goldfish while intensively acquiring geometric spatial information to pass through the gates.     

Induction of diploid gynogenesis in turbot <Emphasis Type="Italic">Scophthalmus maximus</Emphasis> with left-eyed flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis> sperm

Turbot Scophthalmus maximus exhibits sexually dimorphic growth, with females growing faster and reaching larger adult sizes than males. Thus, development of techniques for preferentially producing females is necessary to optimize production of these species. In this paper, gynogenetic diploids of turbot were induced by activating egg development with ultraviolet (UV)-irradiated left-eyed flounder Paralichthys olivaceus sperm combined with cold shock to prevent extrusion of the second polar body. The results of UV irradiation experiments showed that survival, motility, and duration of activity of P. olivaceus sperm generally decreased with increase in UV dose. The typical Hertwig’s effect was observed after fertilized turbot eggs with UV-irradiated P. olivaceus sperm and the optimal UV dose for gynogenetic haploid production was 36,000 erg mm⁻². At 15°C, appropriate timing of cold shock for retention of the second polar body in turbot eggs was at 6 min after fertilization. Results of different combinations of two shock temperatures (1 or 3°C) and four shock durations (15, 25, 35 or 45 min) at 6 min after fertilization demonstrated that shock of 25 min at 1°C gave the highest production of diploid gynogens (39.58% relative to its diploid control). The results of this study reveal that the use of UV-irradiated P. olivaceus sperm for activation of turbot eggs and cold shock for polar body retention is an effective method to produce gynogenetic offspring.