Photoperiodic manipulations of the reproductive cycle of sea bass (Dicentrarchus labrax) and their effects on gonadal development, and plasma 17ß-estradiol and vitellogenin levels

The annual profile of plasma vitellogenin (VTG) and 17ß-estradiol (E₂) levels, as well as gonadal development and spawning characteristics were investigated in captive female sea bass (Dicentrarchus labrax). Endocrine and gonadal changes were studied in fish reared under natural conditions or exposed to manipulated photothermal cycles. In natural conditions of photoperiod and temperature, sea bass spawned from February through March (East coast of Spain, 40°N 0°E). One or two months of constant long-days (15L/9D) in a constant short-day photoperiod regime (9L/15D) all-year-round, given early in the year (March and March–April), advanced spawning by 3 months. The same treatment applied later in the year (September–October) delayed spawning by 1 month, compared to controls.In all groups, changes in plasma VTG levels were correlated with E₂ levels, oocyte growth and spawning time. In control females, VTG was low (<100 ng ml^-1) during the summer, until its first surge in plasma 4 months before the beginning of spawning. The VTG (3.1 ± 0.3 mg ml^-1) and E₂ (4.1 ± 0.5 ng ml^-1) levels showed a single annual peak during late vitellogenesis, the time of the highest proportion of vitellogenic oocytes in the ovary. Constant high levels of VTG (1–1.4 mg ml^-1) and E₂ (1.6–1.9ng ml^-1) were maintained during the entire spawning time, together with the presence of vitellogenic oocytes, suggesting the existence of several waves of oocyte growth in the ovary and thus, several spawns per female. Endocrine profiles and oocyte development in fish exposed to constant photoperiods were similar to controls, but were shifted in time in relation to the displacement of the spawning time. In the fish showing advanced spawns, the duration of the gametogenic proces was compressed when compared to controls. The differences observed in the evolution of the reproductive-related factors in the advanced groups, which were exposed to a reduction in temperature to 15°C, suggest an influence of the temperature in the early stages of the reproductive cycle in sea bass.     

The Effect of Photoperiod on Sexual Maturation,Appetite and Growth in Wild Atlantic Cod (Gadus morhua L.)

An experiment was conducted to examine (a) the effects of photoperiod on timing of sexual maturation (b) the relationship between plasma steroid levels, appetite and growth in male and female Atlantic cod (Gadus morhua L.). Wild caught Norwegian coastal cod were subjected to either a 6L/18D photoperiod typical of January at 60° N-(Short day group) or a simulated natural photoperiod (Normal day group) from June 2000 until spawning started. Appetite of individual fish were measured twice weekly, while weight, length and plasma levels of the sex steroids testosterone (T), 11 keto-testosterone (11-KT) and estradiol-17β (E2) were monitored bimonthly. Cod in the Short day group matured 3 months ahead of the cod in the Normal day group and started spawning in early November. Appetite decreased in both sexes 2–3 months prior to spawning in both groups, but this reduction was stronger among males. In both sexes, length growth was reduced concurrently with the appetite loss. Overall, females had significantly higher somatic growth, put relatively less energy into length growth and had developed larger livers compared to males at the time of spawning in the Short day group. Plasma steroid levels increased in both groups throughout the experiment, reaching peak levels of ca 10 ng ml⁻¹ (T) and 15–20 ng ml⁻¹ (11-KT) in males, and 1.5–2 ng ml⁻¹ (T) and 12–18 ng ml⁻¹ (E2) in females at the onset of spawning. Steroid levels increased more rapidly among Short day cod verifying the earlier onset of maturation. These results confirm that photoperiod is a major cue to maturation in cod and imply that the high cost of spawning for females incur differences in appetite between the sexes.     

Annual cycle of plasma lipids in captive reared striped bass: effects of environmental conditions and reproductive cycle

Protein, lipid and fatty acid concentrations in plasma of eight male and eight female six year old captive striped bass (Morone saxatilis) were monitored monthly over the course of two reproductive cycles as part of an effort to investigate the time course of lipid class mobilization and subsequent deposition in gonads. Total protein levels (44.2 ± 0.67 mg ml^–1, range 20.6–86.4) showed seasonal fluctuation, but did not vary with sex. Total lipid concentrations in the plasma of both males (17.7 ± 0.61 mg ml^–1) and females (14.1 ± 0.64 mg ml^–1) showed seasonal fluctuations with the lowest levels in late Spring during spawning. Plasma lipids in females were significantly (p < 0.0001) lower than those of the males except during early ovarian secondary growth. Analysis of the lipid class composition revealed that the decrease in plasma lipid concentrations in females prior to spawning is primarily due to a decrease of up to 50% in the phospholipid content of the plasma relative to males. Striped bass vitellogenin was found to contain approximately 20% lipid by weight with nearly 80% of the lipid being phosphatidyl choline (PC). Vitellogenin levels previously measured in these individuals were highest during the winter months and decreased in the 2 months prior to spawning. Although mature striped bass oocytes are rich in wax esters, no wax esters or fatty alcohols were found in the plasma. Analysis of the fatty acyl composition of separated lipid classes suggests that PC is the primary carrier of the essential (n=3) fatty acids 22:6 (DHA) and 20:5 (EPA) to the gonads. Fatty acid analysis of the lipids associated with vitellogenin revealed that levels of essential fatty acids were low relative to vitellogenin-borne lipids from other teleosts. The lack of detectable wax esters in plasma lipids of female striped bass suggests that these abundant lipids in mature eggs are synthesized within the oocytes.     

Effect of triiodothyronine on the growth and survival of larval striped bass (Morone saxatilis)

This study was carried out to test the effect of triiodothyronine (T₃) on the growth and survival of larval striped bass (Morone saxatilis). Growth and survival of striped bass held in 5 ppt seawater and treated with various doses of T₃ were measured beginning at 5 and 16 days after hatching. Body content of T₃ was measured by radioimmunoassay. T₃ dissolved in the 5 ppt seawater was taken up by larval striped bass in a dose-dependent manner, and affected the growth and survival of the fish. At 5 days after hatching, T₃ at 100 ng ml^–1 and 50 ng ml^–1 retarded the growth of larval striped bass and caused a lower survival rate than T₃ at 25 ng ml^–1 or the control treatment. At 16 days after hatching, T₃ at 100 ng ml^–1 retarded the growth of larval fish and caused a higher mortality. T₃ at 10 ng ml^–1 and 1 ng ml^–1 did not show any effect on either survival or growth. Body content of T₃ returns to control levels within days following end of treatment. The results indicate that exogenous T₃ can be detrimental to the growth and survival of larval striped bass.     

Sea bream culture in Egypt; status, constraints and potential

Marine fish farming in Egypt began in 1976 with the culture of gilthead sea bream, (Sparus aurata) as this fish was notably adaptable to brackish and marine pond conditions. Today, marine fish and shrimp farms amount to about 19,000 ha, out of which 42% is already in production while the rest, i.e., 58% is still under construction. In 1997, cultured gilthead sea bream production of 2,250 tons made up 3% of the 75,000 tons total aquaculture catch. In polyculture, usually with the grey mullet and sea bass, gilthead sea bream contributed 440 kg ha^–1 to the total yield of 1,700 kg ha^–1 (26%) over a period of 16 months. For the same period, the yield of monoculture ponds averaged 100 kg ha^–1, while in marine cages, yields ranged from 4–10 kg m³. In 1996–1997, fry of 0.25–1 g and fingerlings 1–10 g with a total of 3 million, were collected from the wild and 1 million fry were produced in the three marine hatcheries out of the four existing ones. The development of sea bream culture in Egypt is now severely inhibited by the shortage of seeds and adequate feeds. Exports of both sea bream and sea bass, during 1994–1996 averaged 1,300 tons per year.     

Diel changes in plasma arginine vasotocin, isotocin, and melatonin in rainbow trout (Oncorhynchus mykiss)

The diel changes in plasma AVT, IT and Mel in rainbow trout (Oncorhynchus mykiss) were studied to assess potential relationships. Blood was sampled at 05:00, 11:00, 16:00, 22:30 and 05:00 in freshwater-adapted fish and at 22:30 in brackish water-adapted fish maintained under natural photoperiod. A few of the FW-acclimated fish were assigned to one of two experimental groups and adapted to DD or LL lighting regimes. Blood samples were taken at 11:00 and 22:30. Hormones were extracted from plasma by solid phase extraction and determined by high-performance liquid chromatography. Marked diel variations in AVT and Mel were detected in fish maintained under natural photoperiod. Plasma AVT (fmol ml^–1) increased during the light to reach the maximal level at the end of that phase (261.7±23.1). Thereafter, AVT concentration decreased and became minimal at 05:00 (68.9±11.5) 3 h before the sunrise. Plasma Mel (pmol ml^–1) increased between 16:00 and 22:30 when a peak value was reached (1204.0±55.5). Thereafter, Mel levels decreased and were minimal after the onset of the light phase (242.8±37.0). IT levels displayed no significant diel changes. Linear regression analysis indicated the negative correlation between plasma Mel and AVT for five collecting times of the daily 24 h cycle in freshwater fish and at 22:30 in brackish water fish. A similar correlation occurred at 11:00 in the DD group and at 22:30 in the LL group. To elucidate the character of the Mel-AVT relationships further studies are required.     

Prothrombin time, activated partial thromboplastin time and fibrinogen values in Mediterranean marine teleosts

Coagulation profiles for marine fish were determined with the standard global screening assays, the prothrombin time (PT) and activated partial thromboplastin time (APTT) using homologous tissue thromboplastin and phospholipid extracts to investigate species-specificity of brain thromboplastins, and to establish a base-line coagulation profile for marine fish of economical importance for Mediterranean aquaculture (sea bream, Sparus aurata, sea bass, Dicentrarchus labrax, red porgy, Pagrus pagrus, common dentex, Dentex dentex). Sea bass and sea bream displayed the shortest PT (9.0–15.0 s) and APTT (22.1–35.8 s) in comparison to the other species (PT: 24.8–37.2 s, APTT: 39.6–66.4 s). Red porgy displayed the lowest fibrinogen value (1.2–2.3 g l^–1), while no differences were found among the other species (2.2–2.8 g l^–1). It is concluded that the interaction of the tissue factor and plasma clotting factor is relatively species specific and that the use of homologous brain extracts is preferable for haemostasis study in marine fish.     

Disruption of gonadal maturation in cultured Senegalese sole Solea senegalensis Kaup by continuous light and/or constant temperature regimes

A major problem in the development of Senegalese sole Solea senegalensis intensive culture is the poor control on reproduction, in part due to the lack of knowledge on the precise role of photoperiod and temperature. Thus, gonadal maturation was evaluated by assessing the sequential changes in plasma levels of 17β-estradiol (E₂), 11-ketotestosterone (11-KT), and testosterone (T) in both female and male cultured Senegalese sole (F1 generation) exposed to various combinations of constant or naturally-fluctuating daylength and water temperature. Under natural photoperiod (NP; 36° N), exposure to constant temperature (t0; 18-20 °C) disrupted gonadal development, as indicated by a lower incidence (in comparison with naturally-fluctuating water temperature; 14-24 °C) of females at advanced maturation (from February to April: 12 vs. 33%) and running males (from February to May: 46% vs. 57%), and the reduced mean (± S.E.M.) sex steroid plasma levels (female peak E₂ levels: 2.9 ± 0.28 vs. 1.8 ± 0.3 ng ml^− 1; male peak T levels: 1.5 ± 0.14 vs. 0.9 ± 0.06 ng ml^− 1). Therefore, the onset and progression of gonadal development in this species seem to be strongly (“proximally”) influenced by fluctuating water temperature. When compared to NP and t0, exposure to continuous light (LL) under t0 significantly reduced steroid production (female peak E₂ levels: 1.8 ± 0.28 vs. 0.5 ± 0.05 ng ml^− 1; male peak 11-KT levels: 9.4 ± 1.06 vs. 5.4 ± 1.33 ng ml^− 1) and subsequently gonadal development (lower proportions of females at intermediate [46 vs. 6%] and advanced maturation [12 vs. 0%] from February to April and of RM [46 vs. 33%] from February to May). Thus, the seasonal changes of daylength would be crucial for normal gonadal development, being its cueing effects of higher magnitude than those of water temperature. The present report constitutes the first systematic study focused on the environmental control of reproductive events in Senegalese sole.     

A new technique of feeding, repeated sampling of blood and continuous collection of urine in white sturgeon

A new technique combining oesophageal intubation, dorsal aorta cannulation, and urinary catheterization was developed to monitor concentration of nutrients in the blood circulation and their metabolites in the urine of 1–2 kg white sturgeon, Acipenser transmontanus. Three experiments were conducted to assess the technique based on: (1) quantitative delivery of nutrients; (2) stress levels post-operation; and (3) monitoring of nutrients in the blood and metabolites in the urine. In Experiment 1, recovery of intubated Cr₂O₃ was 105±5% (mean ± SEM, n=3) 3 h after intubation. In Experiment 2, plasma cortisol and glucose concentrations returned to respective basal levels of 8.2±1.8 ng ml^–1 and 74± mg dl^–1 (n=9) 48 h post-operation. In Experiment 3, sturgeon intubated with 1 g kg^–1 body weight of glucose or dextrin at 48 h post-operation showed a significantly different (p<0.05) peak plasma glucose level of 139±4 and 100±5 mg dl^–1 (n=5), respectively, at 4 h. Urinary glucose excretion was 1.8±0.9 mg kg^–1 h^–1 (n=5) 4–8 h after intubation with glucose. Our results show that the new technique allows quantitative delivery of nutrients, repeated sampling of blood, and continuous collection of urine in white sturgeon with minimum stress.     

Production of melatonin by the gilthead sea bream pineal: anin vivo andin vitro study

This study deals with the variations of plasma melatonin content and ofin vitro pineal melatonin secretion, in one year-old male sea bream. Plasma melatonin levels varied from almost undetectable levels during the day to 150–200 pg ml^–1 during the night, in animals maintained under a light/dark (12L/12D) cycle. The duration of the nocturnal surge corresponded to the duration of the night. A similar pattern of melatonin secretion was obtainedin vitro, with pineals cultured under a 24h L/D cycle. When the photoperiod cycle was advanced by 8h, the first nocturnal peak in melatonin secretion was more acute compared to the profile described above, but the rhythmic variations were immediately synchronized to the new L/D conditions. Under constant darkness (D/D), circadian variations in melatonin release were observed for no more than two 24h cycles. No melatonin was detected in the culture media from retinas cultured under L/D or under D/D. These results strongly suggest that the pineal is the major source of plasma melatonin in the gilthead sea bream, and that the 24h photoperiod cycle is a powerful synchronizer of the rhythmic production of melatonin by this organ.     

Nitrogenous waste excretion and accumulation of urea and ammonia inChalcalburnus tarichi (Cyprinidae), endemic to the extremely alkaline Lake Van (Eastern Turkey)

The endemic, anadromous cyprinidChalcalburnus tarichi is the only fish species known to occur in alkaline Lake Van (Eastern Anatolia, Turkey). EightC. tarichi were maintained individually in Lake Van water (17 – 19°C; pH 9.8; 153 mEq·I^–1 total alkalinity; 22 total salinity) and tank water samples analyzed for 24 h in 2 to 4 h intervals. At zero time, < 1µM ammonia was present and urea was undetectable in the tank water; at 24 h, total ammonia and urea made up 114±32 and 35±25µM, respectively. Over the experimental period, ammonia-N and urea-N excretion averaged 1041±494 and 607±169moles·kg^–1 fish·h^–1, respectively. The extent of urea excretion was highly variable between specimens. Uric acid excretion was not detectable.Urea was present at high concentrations in all tissues and plasma (25 – 35moles·g^–1·ml^–1) of freshly caughtC. tarichi; total ammonia content of the tissues was by a factor of 1.9 (liver) to 3.0 (brain) lower. High arginase activity (2.4±0.2 U·min^–1·g^–1) was detected in the liver ofC. tarichi but ornithine carbamoylphosphate transferase, a key enzyme of the ornithine-urea-cycle, was absent. Ureagenesis is likely through degradation of arginine and/or uricolysis. High glutamine synthetase activity (11±0.6 U·min^–1·g^–1) and low ammonia content in brain suggest that, like other teleosts,C. tarichi has an efficient ammonia detoxification in the brain, but in no other tissue.Nitrogenous waste excretion at alkaline pH is discussed. The ability ofC. tarichi to excrete high levels of ammonia at extremely alkaline pH is unique among teleosts studied so far. The mechanism of ammonia excretion under Lake Van conditions remains to be elucidated.     

Potassium flux in juvenile striped bass (Morone saxatilis): influence of external concentrations of sodium chloride and calcium

Whole-body influx and efflux of K⁺ were determined for 25-day-old striped bass,Morone saxatilis, in conditions that simulated harvesting fish from ponds. During the first 5h in fresh water with combined high NaCl (80 mM) and low Ca²⁺ (0.12 mM) concentrations, a combination that is acutely lethal to this age of striped bass, K⁺ influx for fish in 0.07 mM K⁺ was 21±1.7 (SEM) compared to 3.4±0.33 nmol g^–1 h^–1 for fish in water with low Na⁺ (0.25 mM) or high Ca²⁺ (2.5 mM) concentrations. Influx of K⁺ was inhibited during the first few hours after fish were placed in flux chambers. Potassium efflux as percentage of⁴²K lost per hour was two-fold higher from fish in the high Na-low Ca treatment compared to fish in low concentrations of Na⁺ or high concentrations of Ca²⁺. Potassium efflux was probably much greater than influx, but exact values for efflux could not be calculated from the data available. Survival of fish in water with high Na-low Ca was not increased by addition of KCl to the water, indicating that the net loss of K⁺ was probably not the cause of death.     

Stress-induced changes in the affinity and abundance of cytosolic cortisol-binding sites in the liver of rainbow trout,Oncorhynchus mykiss (Walbaum), are not accompanied by changes in measurable nuclear binding

Plasma cortisol levels and the number (N_max) and affinity (K_d) of specific hepatic cortisol-binding sites were determined in rainbow trout subjected to chronic confinement stress for 14 days. Confinement significantly elevated plasma cortisol levels to 47.3 ± 13.5 ng ml^–1 within 24h and although levels declined to 8.0 ± 3.0 ng ml^–1 after 14 days, they were significantly higher throughout than levels in unstressed control fish (< 2.0 ng ml^–1). There was a 60% reduction in cytosolic N_max in stressed fish during the first 24h of confinement (35.8 ± 7.9 cf. 129.0 ± 15.2 fmol mg^–1 protein), a decline which was sustained at 7 days after the onset of stress but, although numbers of binding sites in the liver of stressed fish were still lower than in unstressed fish, the difference was no longer significant after 14 days of confinement. There was an accompanying significant rise in the K_d of cortisol binding in stressed fish during confinement, from 4.0 ± 0.6 nM at time 0 to 8.4 ± 0.8 nM after 24 h confinement. This increment in K_d was sustained at a level significantly higher than in control fish throughout the 14 day confinement period, despite marked reductions in cortisol levels and increases in N_max in stressed fish. Throughout the study, specific binding of cortisol could not be consistently detected in high-salt nuclear extracts from stressed or unstressed fish, suggesting either that high-affinity binding sites for cortisol were absent from these preparations, that receptors were present but unable to interact with ligand because they were occupied, or that receptors were present but not being extracted. These possibilities were investigated using a range of extraction procedures, by varying the temperature of incubation, by employing dexamethasone as ligand and by examining binding in purified, intact, nuclei. Estradiol was employed as a methodological control throughout and substantial amounts of specific estradiol binding were detected in all compartments and preparations. Specific cortisol-binding sites were detected in intact nuclei of both stressed and unstressed fish, at levels an order of magnitude lower than estradiol binding in the same preparations. These data demonstrate that activation of the pituitary-interrenal axis leads to significant changes in the nature of target-tissue binding of cortisol in rainbow trout, and reveal a clear difference in the subcellular distribution of binding-sites for estradiol and cortisol, which reflects the situation in mammalian tissues.     

Dehydration is more effective for the control of embryonic development and larval hatching of Diplectanum aequans (Monogenea, Diplectanidae) than formalin and trichlorphon

Embryonic development and larval hatching of the monogenean Diplectanum aequans, gill parasite of sea bass Dicentrarchus labrax, was studied in relation to different prophylactic treatments. Groups of eggs of D. aequans were submitted to different in vitro treatments: formalin (300 and 100 L L^–1 per 1 hour), Neguvon^® (trichlorphon 0.2 mg L^–1 per 48 hours) and dehydration for 4 and 8 hours. Percentages of hatched larvae, aborted larvae and undeveloped embryos were estimated in comparison with the control group. Results showed that 300 L L^–1 formalin and dehydration treatments were able to reduce larval hatching significantly, while Neguvon^® and 100 L L^–1 formalin treatments had no effect.     

A comparative study of some hematological features in young reared sturgeons (Acipenser persicus and Huso huso)

The physiological condition in fishes required for their selection as broodfish can be determined by hematological studies. The present study aims at examining the hematological indices in juvenile sturgeons at different stages of growth. The main blood features, such as total albumin content in blood, hemoglobin concentration and the erythrocyte and leukocyte count, were measured in a total of 54 specimens of 1-, 2- and 6-year old sturgeon (Persian sturgeon Acipenser persicus and beluga Huso huso), artificially produced at the Shahid Dr. Beheshti Fish Breeding and Rearing Center located in the Gilan Province.In the Persian sturgeon specimens studied, the total albumin concentration in the blood increased from 26.05 to 52.50 g l^–1, the hemoglobin concentration increased from 42.2 to 65.8 g l^–1, the erythrocyte count increased from 240 to 452.5 thousand cell ml^–1 and the leukocyte count increased from 13.43 to 46.48 thousand cells ml^–1. The total leukocyte was composed of 73.25 to 82.70% lymphocytes, 12.3 to 20% neutrophils, 2.25 to 6.50% eosinophils and 0.2 to 2.5% monocytes.In the beluga specimens studied, the total albumin concentration in the blood increased from 20.05 to 42.20 g l^–1, the hemoglobin concentration increased from 46.3 to 78.6 g l^–1, the erythrocyte count increased from 495 to 735 thousand cells ml^–1 and the leukocyte count increased from 31.6 to 57.7 thousand cells ml^–1. The total leukocyte was composed of 54.5 to 67.5% lymphocytes, 22.63 to 33.86% neutrophils, 6.60 to 13.7% eosinophils and 0.6 to 2.25% monocytes.This research reveals the trends of some hematological features during the growth and development of sturgeons in the South Caspian Sea, reared in artificial conditions. It shows that with the increase in age, the hematological indices also increased.     

Effect of different schedules for broodstock exchange on the seed production of Nile tilapia Oreochromis niloticus (L.) in freshwater

This study compares the effect of three broodstock exchange treatments (exchange every month, two months and three months) versus no-exchange control treatment on year-round seed production and spawning synchrony of Nile tilapia Oreochromis niloticus stocked in 0.4 m³ fiberglass tanks within a recirculating water system. Adult males and females with mean body weights of 161.7 g and 127.6 g, respectively, were stocked at a rate of 4 fish/tank with a male to female sex ratio of 1:3. Water temperature at 29 ± 1.0 °C and photoperiod of 18 h/day were maintained throughout the experiment. The results showed that broodstock in the exchange treatments had higher seed production and spawning synchrony compared with those in the no-exchange treatment. The exchange treatments resulted in 70.5 to 97% increase in seeds kg female^–1 day^–1. The different spawning parameters of broodstock in the two- and three-month exchange treatments were higher than the one-month exchange treatment. Results suggest that broodstock exchange strategy has a long-term improvement effect on seed production and spawning synchrony, and exchanging all breeders every three months is more profitable in terms of better utilization of breeders and in reducing both the holding/conditioning facilities for broodstock and the labor cost.     

Incidence and prevalence of Diplectanum aequans and its influence on the fitness of juvenile sea bass (Dicentrarchus labrax) in the Black Sea

High rates of unexplained mortalities (up to 70%) are anticipated in the cultured juvenile sea bass (Dicentrarchus labrax) in farms in the eastern Black Sea at least for the last 8 years. Diplectanum aequans, often blamed for the observed mortalities, is the only prevalent parasite impacting the health of sea bass in the brackish water of the Black Sea. To investigate the seasonal fluctuation of D. aequans prevalence, mean intensities and the potential effects of D. aequans on the fitness of sea bass, eight cages from one farm (Farm A) and four cages at another distinct site (Farm B) were surveyed monthly from May to October of 2008. Six hundred and sixty‐one juvenile sea bass from Farm A and 236 from Farm B were individually examined for the presence and intensity of the parasite. The prevalence of D. aequans in cultured sea bass (96.3 ± 11.11; Mean ± SD) ranged from 66.7% to 100% in June, the beginning of 6‐month growing season. The mean intensity was 6.08 ± 2.19, significantly higher than that (2.74 ± 0.87; P < 0.05) in December (P < 0.05), the end of the growing season. Fish with low fitness had significantly higher number of parasite than the fish with higher fitness (P < 0.005). Although D. aequans negatively influenced host's condition factor, the impact was not enough to lead mortalities in the cultured sea bass in the Black Sea.     

Off-Season Spawning of Eurasian Perch Perca Fluviatilis

This study was designed to determine the effects of the selection for body weight and of two final warming durations during an out-of-season reproductive cycle on gonad recrudescence, plasma steroid levels and spawning in Eurasian perch Perca fluviatilis males and females. Two different bodyweight classes (200–250 and 350–400 g) and two different warming periods (1 or 2 months) were studied.At the end of the chilling period, a significant effect of the selection for body weight was observed on gonad development whereas fish were the same age. According to higher levels in testosterone (12 ng mL^–1), a better development was recorded for smaller breeders (P < 0.05). A short water warming period up to 14°C (1 month v.s. 2 months) allowed higher rates of females with completed gonadogenesis (33–38% v.s. 7–8%) and spawning (31–33% v.s. 0%) (P < 0.05). Spontaneous out-of-season spawning (17 ribbons) and larvae were obtained. Relative fecundity was about 100 eggs g^–1 of female body weight. Five ribbons were partly fertilized with a rate ranging from 5 to 90%. These studies have shown that delayed spawning in Eurasian perch can be obtained by a temperature manipulation, but results are not sufficient because of low stimulation rate of females, low spawning rates and high heteroge-neity of fertilization rates. The effects of other environmental factors like photoperiod should be tested in further experiment to improve the protocol inducing out-of-season spawning production.     

Tilapia wild spawning control through predator fishes: Israelitrial with red-drum and hybrid bass

Tilapia wild spawning is a nuisance in warm freshwater aquaculture growout ponds. To cope with this problem two experiments were carried out with predatory fish that do not reproduce in fresh water. One experiment tested the capacity of hybrid bass (Morone saxatilis × M. chrysops) and red-drum (Sciaenops ocellatus) as predators of wild spawning of hybrid tilapia (Oreochromis niloticus × O.aureus), and the other compared predation effectiveness of red-drum of different sizes and stocking densities.Both hybrid bass and red-drum effectively reduced tilapia wild spawning and improved by 15–20% tilapia performance and food conversion ratio. These effects were obtained stocking small red-drum (20 g) or large red-drum (60–80 g) or bass (135 g) at stocking densities of 500–1000 predators/ha, together with 15000 tilapia/ha of 65–75 g. Hybrid bass stocked at 750/ha and large red-drum stocked at 500/ha presented over 90% survival. Red-drum at higher stocking density and/or lower stocking weight presented reduced survival (40–60%). Red-drum of all examined stocking weights presented better growth rates when stocked at 500/ha than at higher densities.     

New perspective to control of tenacibaculosis in sea bass Dicentrarchus labrax L

This work was carried out to investigate the effects of injection of Tenacibaculum maritimum formalin‐killed cells (FKC), extracellular products (ECPs) and crude lipopolysaccharide (LPS) as well as 1% feed supplements of oil extracts of Echinacea purpurea and Origanum vulgare on sea bass immunity improvement to the favour of T. maritimum experimental infection control after 4 weeks of the experiment. Tenacibaculum maritimum isolated from naturally infected sea bass showed brown to yellowish‐brown lesions (sores) on gills, skin and/or fins and identified by different biochemical methods and polymerase chain reaction technique. Immune parameters namely, total protein, globulin and lysozyme activity, as well as the relative level of protection were improved by T. maritimum (FKC), (LPS), (ECPs), O. vulgare and E. purpurea, respectively compared with control. Histopathological examination of T. maritimum naturally infected sea bass indicated many pathological changes in gill, skin and musculatures. Present study could be concluded that application of T. maritimum (FKC), (LPS), (ECPs), O. vulgare or E. purpurea improved sea bass immunity to the favour of disease resistance against T. maritimum. Further investigations on the combination between the previous control methods and the vaccine application methods will be needed.