Effects of yeast hydrolysate supplementation in low-fish meal diets for pikeperch.

Plant proteins have been increasingly used as sustainable substitutes for fish meal (FM) in aquafeeds; however, their high inclusion level compromises fish performance. The objective of this study was to examine whether yeast hydrolysate (YH) supplementation can improve the utilisation of high soybean meal (SM) diet and ameliorate its potential deteriorating impacts in pikeperch (Sander lucioperca). A basal diet was formulated using 44% FM, and four additional diets were produced by replacing 30 or 60% of FM with SM with or without the addition of 2% YH (FM, SM30, SM60, SM30 + YH, and SM60 + YH diets). Each diet was fed to three groups of fish (35.3 ± 0.10 g, 150 fish per group) to visual satiety four times daily for 70 days. Fish growth was not impacted by FM replacement level or YH application. However, SM60 group exhibited markedly higher feed conversion ratio and lower survival rate than those fed the FM- and YH-supplemented diets (P < 0.05). The highest and the lowest protein efficiency ratio values were obtained for the SM30 + YH and SM60 groups, respectively. Whole-body lipid content decreased in SM60 and SM60 + YH groups, and muscle lipid decreased in all the replacement groups. Serum triglyceride and glucose concentrations tended to decrease as FM replacement level increased. The highest alanine aminotransferase, aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) activities were detected in the SM60 group, and YH addition significantly decreased the AST and LDH activities. Serum lysozyme activity decreased in SM30, SM60 and SM60 + YH groups. Serum myeloperoxidase and antiprotease activities decreased in SM60 group, and YH supplementation improved their activities. No effects of diets were observed on serum antioxidant parameters such as catalase activity and malondialdehyde concentration, and gut morphological indices. Number of goblet cells in midgut decreased by increasing the SM inclusion level and a slight improvement was observed by YH application. These findings suggest that YH supplementation has the potential to support the replacement of up to 60% FM with defatted SM in pikeperch feed without deteriorating growth, feed utilisation, and survival rate. Further, YH incorporation mitigated the damaging impacts of high SM diet on liver function and non-specific immune response.

Optimization of short-term storage of pikeperch semen: an applicable approach.

Contamination of semen with urine and asynchronous maturation of males and females are main obstacles in artificial reproduction of pikeperch Sander lucioperca. The objective of this study was to overcome these obstacles using optimization of a procedure for short-term storage of pikeperch semen at 4°C using two immobilizing media (IM): (a) IM1, 180mM NaCl, 2.68mM KCl, 1.36mM CaCl2⋅2H2O and 2.38mM NaHCO3, 343mOsm/kg; and (b) IM2, 200mM NaCl, 2.68mM KCl, 1.36mM CaCl2⋅2H2O and 2.38mM NaHCO3, 381mOsm/kg. Undiluted sperm was used as the control. At 6h poststorage, there were no substantial changes in spermatozoa motility and velocity at 30s postactivation in all groups. Over 48h of storage, the highest spermatozoa motility and velocity were obtained in sperm diluted in IM2 compared to the other groups. IM2 could maintain a significantly higher ATP content of diluted sperm than IM1 and undiluted treatment for 2days. Similarly, the highest values of eyeing and hatching rates were observed in sperm diluted in IM2 compared to sperm in the other studied groups. It can be concluded that the obtained result is a novel and applicable approach to maintain semen quality of pikeperch during short-term storage, suggesting IM2 as a promising medium for short-term storage. The present study also opens possibilities for ensuring a reliable source of semen as a convenient approach for increasing genetic diversity in hatcheries.

D1, but not D2, dopamine receptor regulates steroid levels during the final stages of pikeperch gametogenesis.

In pikeperch, Sander lucioperca, aquaculture hormonal treatment is usually applied to synchronize ovulation. However, the effect of dopamine (DA) receptor antagonists, in particular those blocking the D1 DA receptors, remains unknown. Thus, the aim of the present study was to investigate and compare the effects of D1 and D2 DA receptor antagonists on the sex-steroid production and reproductive performance of the species. Two experiments were performed during which mature pikeperch females were injected with different molecules: NaCl 0.9% (negative control) or human chorionic gonadotropin 500 IU/kg (positive control) in both experiments, metoclopramide (a D2 receptor antagonist; 4 mg/kg or 20 mg/kg) or SCH23390 (a D1 receptor antagonist; 0.8 mg/kg or 4 mg/kg) alone (experiment 1) or in combination with a salmon gonadotropin-releasing hormone analogue (sGnRHa at 25 µg/kg; experiment 2). In experiment 2, fish were also injected with sGnRHa (25 µg/kg) as positive control. Samplings of oocytes and blood were performed on the day of injection and after 24 h (both experiments), after 48 h (experiment 2) and at the time of ovulation (both experiments). In non-ovulating fish, samplings were performed 7 days (experiment 1) or 14 days (experiment 2) after injection. In experiment 2, various zootechnical parameters of fertilized eggs were recorded (survival, hatching and malformation rates). The two antagonists alone were ineffective in inducing the final stages and regulating sex-steroid (testosterone, 11 ketotestosterone, 17ß estradiol and 17,20ß-dihydroxy-4-pregnen-3-one) production. When administered with sGnRHa, both SCH23390 and metoclopramide induced the final stages. However, only SCH23390 stimulated testosterone (4 mg/kg) and 17ß estradiol (0.8 mg/kg) production compared with sGnRHa alone. None of the treatments affected the survival, hatching or malformation rates. This is the first report suggesting that in pikeperch the D1, but not the D2, DA receptor antagonist would be involved in the testosterone and 17ß estradiol production as a potentiator of the sGnRHa effect.

Rearing conditions and life history influence the progress of gametogenesis and reproduction performances in pikeperch males and females.

Pikeperch (Sander lucioperca) is a highly valuable fish in Europe. However, development of aquaculture of pikeperch is highly limited due to seasonality of production. This can be overcome by the controlled reproduction of domesticated fish. The first steps of domestication process may induce changes at anatomical, physiological and molecular levels, thereby affecting a variety of biological functions. While there is abundant literature on their effects on stress and growth for example, these effects on reproduction received limited attention notably in pikeperch, a promising candidate for the development of aquaculture. To answer the question of this life-history effect on pikeperch's reproduction, we compared two groups (weight: 1 kg) originated from Czech Republic and with the same domestication level (F0). The first group was a recirculating aquatic system cultured one (2 years, previously fed with artificial diet, never exposed to natural changes in temperature/photoperiod conditions) and the second one was a pond cultured group (3 to 4 years, bred under natural feeding and temperature/photoperiod). The wild group successfully spawned, while the farmed one did not spawn at all. During the program, gonadosomatic indexes of both males and females were significantly higher for the wild fish, as well as the sexual steroids. Gene expression analysis revealed significantly lower LH transcript levels at the pituitary level for the farmed females and lower FSH transcript levels at the pituitary level for the males. In conclusion this study showed that the previous rearing conditions (e.g. culture system, age, diet, etc.) alter the further progress of gametogenesis and the reproductive performances in response to controlled photothermal program for both sexes in pikeperch.

Development, and effect of water temperature on development rate, of pikeperch Sander lucioperca embryos.

Knowledge of embryo development is essential to the application of reproductive biotechnology in aquaculture, including for pikeperch Sander lucioperca. We describe pikeperch embryo development and demonstrated effects of temperature on the duration of embryogenesis. Developmental stages in embryos incubated at 15 °C were identified as zygote, 0-1.5 h post-fertilization (hpf); cleavage, 2.5-7.5 hpf; blastula, 9-18.75 hpf; gastrula, 21-39, hpf; segmentation, 45-105 hpf; and hatching, 125-197 hpf. Additional groups of eggs were fertilized and incubated at 10, 15, 20, and 25 °C to document stages of development, development rate, and survival. The optimal fertilization and incubation temperature was shown to be 15 °C, with the highest fertilization, survival, and hatching rates. Embryo development was slower at 10 °C, with 45% of fertilized embryos surviving to hatching. Development was accelerated at 20 °C, and resulted in a 56% survival rate of fertilized embryos. At 25 °C, embryos did not develop to the blastula stage. Pikeperch could be a valuable percid model for research in which flexible incubation temperatures is required.

In vitro storage of unfertilized eggs of the Eurasian perch and its effect on egg viability rates and the occurrence of larval malformations.

Ova ageing is the most important factor affecting fish egg quality after ovulation. Long-term storage of fish ova, using cryopreservation and vitrification techniques, has been unsuccessful to date. Instead, short-term in vitro ova storage has been used successfully and optimized in some cultured fish species. In vitro ova storage can drastically improve mass production of larvae and juveniles in the hatcheries by providing the possibility of the synchronous artificial fertilization for different females. To study how long unfertilized eggs of Eurasian perch (Perca fluviatilis L.) can retain their fertilizing ability after stripping, eggs were stored at temperatures of 4°C, 8°C and 12°C for 72 h post-stripping (HPS). The stored eggs of four female perch were separately fertilized at 0 h (i.e. control eggs fertilized before storage) and at 6-hour intervals during the experimental period of 72 h. The embryos reaching the eyed-egg and hatched-larvae stages, eyed-egg mortality and larval malformation rates were recorded as indices of egg quality. The results indicated that the maximum eyed eggs and hatched larvae (86% and 63%, respectively) were observed for eggs fertilized immediately after stripping, whereas the storage of the eggs at 4°C for 48 HPS decreased the eyed-egg and hatched-larvae rates to 46% and 17%, respectively. The use of a higher storage temperature resulted in a more rapid decrease in egg viability: eyed-egg and hatched-larvae rates of 23% and 9%, respectively, were obtained after 48 HPS storage at 8°C and 2% and 1% for eggs stored at 12°C. Eyed-egg mortality and larval malformation rates were not significantly affected by post-stripping ova ageing for at least up to 36 h. Thereafter, both values increased significantly and were measured to be the highest in the most aged ova. The present study demonstrated that stripped Eurasian perch eggs can be stored for at least 12 h at 4°C to 12°C without a significant reduction in their quality.

Splenic lipidosis in intensively cultured perch, Perca fluviatilis L.

Macroscopically visible lipid deposition varying in size from pinpoint to 8-mm diameter was found in spleens of a population of intensively farmed perch, Perca fluviatilis L. over a 24-month rearing period. Large agglomerates of adipocytes distinguishable from surrounding normal tissue occurred in all individuals with spleen lipidosis. Several affected fish presented total dystrophy of large clusters of hepatocytes. Prevalence of lipidosis was 5.0% at 12 months and 16.6% at 24 months. There was no significant difference between fatty acid profiles of liver or perivisceral fat of perch with and without lipidosis except for linoleic, myristic, γ-linoleic, cis-eicosatrienic, palmitooleic acid. Body weight and hepatosomatic, perivisceral fat and splenosomatic indices were not associated with lipidosis. There was no significant effect of lipidosis on mortality or growth.

Planktonic or non-planktonic food in young-of-the-year European perch Perca fluviatilis in ponds.

Higher biomass especially of some aquatic macrophyte species offered a higher density of phytophilous zoobenthos, but did not increase the proportion of non-planktonic to planktonic prey in young-of-the-year perch Perca fluviatilis. Both abundance and biomass of non-planktonic prey dominated over planktonic prey in the pond with lower biomass of aquatic macrophytes and lower food. Survival of P. fluviatilis was lower (20%) in the pond with lower food than in the other pond (34%), however, specific growth rate (1.3% day(-1) ) and final Fulton's condition factor of P. fluviatilis were similar in both ponds.

PCR detection of the crayfish plague pathogen in narrow-clawed crayfish inhabiting Lake Egirdir in Turkey.

Many populations of the narrow-clawed crayfish Astacus leptodactylus in Turkey, including those inhabiting Lake Egirdir, declined drastically in the mid-1980s due to introduction of crayfish plague Aphanomyces astaci. However, unlike many other localities, there has been some recovery in the A. leptodactylus population inhabiting this lake even though crayfish plague has been suspected to have persisted since then. In support of this, DNA from 5 of 34 healthy-looking crayfish sampled recently from the lake tested positive by both conventional and real-time PCR using species-specific primers targeting the rDNA internal transcribed spacer region, and product sequence analysis confirmed the identification of A. astaci. This complies with other recent reports of coexistence of native European crayfish with this pathogen, and further research is now needed to identify the key mechanisms allowing it.

Sensory and textural attributes and fatty acid profiles of fillets of extensively and intensively farmed Eurasian perch (Percafluviatilis L.).

Sensory attributes, texture and fatty acid profiles of fillets of Eurasian perch (Percafluviatilis L.) reared under two conditions were compared. Perch were reared either in an extensive pond-based (EC) system in polyculture with carp, or intensively cultured (IC) in a recirculation system. Attributes of raw and cooked fillets of marketable perch (120-150g) were compared. No significant differences were found between groups for odour, flavour, aftertaste, or consistency in subjective evaluation of cooked fillets. The texture profile analysis (TPA) showed raw fillets from the EC group to exhibit higher values of hardness, springiness, cohesiveness, and gumminess than the IC group. Fish from the IC group had a lower content of saturated fatty acids (SFA) and polyunsaturated fatty acids (PUFA) and a higher content of monounsaturated fatty acids (MUFA) in comparison to EC perch. The proportion of iso- and anteiso-SFAs was 2.6% in the EC group and 0.75% in the IC group. The content of n-3 PUFA was lower in IC than in EC, while the content of n-6 PUFA was higher in IC than in EC. The ratio of n-3:n-6 PUFA was 1.42 for the IC group and 2.85 for the EC group.

Semen of Perca fluviatilis L.: sperm volume and density, seminal plasma indices and effects of dilution ratio, ions and osmolality on sperm motility.

The objectives of the present study were to characterize sperm volume and density, seminal plasma indices (ionic contents and osmolality) and to study the effects of dilution ratio, ions and osmolality on sperm motility parameters (percentage of motile sperm and sperm velocity) in farmed European perch (Perca fluviatilis L.). The means of sperm volume (ml), sperm density (x10(9)spermml(-1)) and total number of sperm (volumexdensity) per fish were 2.75+/-0.51, 29.19+/-3.15 and 82.19+/-15.26. The seminal plasma osmolality (mOsmkg(-1)), sodium, chloride, potassium and calcium ions concentrations (mM) were measured to be 298.07+/-5.09, 130.97+/-2.19, 106.75+/-2.37, 10.70+/-0.61 and 2.41+/-0.09, respectively. At 15s post-activation of stripped sperm, the percentage of motile sperm (%) and sperm velocity (mums(-1)) were 91.90+/-1.27 and 115.54+/-1.25, respectively, and decreased significantly following sperm activation (P<0.05). The optimal sperm motility was observed when the sperm was prediluted in immobilizing solution (IS) at a ratio 1:50. Prediluted sperm showed the maximum velocity when activated in 2.5mM Ca(2+), 50mM K(+) and sucrose with osmolality 100mOsmkg(-1). Neither Ca(2+) nor K(+) showed a significant effect on the percentage of motile sperm at 15s post-activation. Osmolality higher than 200mOsmkg(-1) significantly decreased the percentage of motile sperm, while osmolality of 300mOsmkg(-1) or above totally suppressed sperm motility.