Production of meiotic gynogenetic and triploid sea bass, Dicentrarchus labrax L.: 1. Performances, maturation and carcass quality

Meiotic gynogenetic and triploid sea bass were produced by pressure shocks according to a previously published protocol. Pressure-treated groups did not survive as well as controls during early development and larval rearing. Performances, sexual maturation and carcass quality were examined over a period of 34–45 months. At the age of 34 months, growth of the gynogenetic fish was comparable to that of the control but inferior in the triploid fish. A predominance of male fish was found within the triploid groups, while diploid and meiotic gynogenetic fish showed equal proportions of the sexes. Gonadal maturation in triploid fish was significantly impaired, particularly in the females that showed rudimentary ovaries. Triploid males exhibited primary maturation but proved to be gametically sterile. Pressure-induced triploids did not grow as well as diploids, but these results might be ascribed to specific on-growing conditions (communal rearing). The performance of gynogenetic sea bass was comparable to that of control. The superiority of diploid fish over their triploid counterparts was confirmed during the final growing period and more clearly so in females. Performances of triploids varied according to their maternal origin. Overall, striking qualitative differences between diploid and triploid fish were found at the age of 34 and 45 months, although the results varied in a gender-specific manner. A strong maternal effect was also observed. The potential advantages of triploid sea bass for aquaculture purposes are discussed.     

Aquaculture performance of triploid barfin flounder Verasper moseri

ABSTRACT:   To evaluate the aquaculture performance of triploid barfin flounder Verasper moseri , the sex ratio, maturation, growth and the relative proportion of body parts were examined. The sex ratio of triploids was similar to diploids under communal rearing conditions, but the proportion of female diploids was higher than that of triploids under separate rearing conditions. The gonadosomatic index of triploid females was very low even during the spawning season, and the ovaries were rudimentary. These results suggest that triploid barfin flounder females were sterile. In addition, triploid males produced a small quantity of milt containing very few spermatozoa with abnormal shapes. Spermatozoa obtained from triploids were aneuploidies. When normal eggs were fertilized with sperm from triploid males, no fry developed. These results suggest that triploid barfin flounder males were functionally sterile. Triploid males grew more slowly than diploid males, and triploid females showed similar or slower growth than diploid females, whether reared separately (23 months) or communally (35 months). The ratios of visceral weight to the edible parts for triploid males were similar to those for diploid males, but ratios for triploid females were higher than for diploid females during the spawning period. In conclusion, a significant improvement of growth was not found in triploid barfin flounders.     

Plasma levels of Gonadotropin-II and gonadal sex steroids in triploid catfish, Heteropneustes fossilis (Bloch)

Triploid fish have under-developed gonads due to altered reproductive endocrinology. Triploids of Indian catfish (H. fossilis) showed significantly reduced plasma levels of gonadotropin (GtH-II), testosterone (T) and estradiol-17 (E₂) than that of diploids throughout the year, except for the resting phase, irrespective of sex. Plasma levels of GtH-II were significantly different (p<0.001) between diploid and triploid fish during preparatory, prespawning and spawning phase. The plasma testosterone contents in triploids were significantly less (p<0.001) than that of diploids, except during the resting phase. Triploid females showed very low titres of estradiol-17 (<1 ng ml^–1) throughout the annual reproductive cycle in contrast to highly fluctuating levels in diploid females. Thus, this study for the first time provides information on reduced levels of GtH-II and sex steroids in plasma of male triploid fish and additional information on species-specific alteration of sex hormones in female triploids.     

Reproductive ability of triploid ornamental (koi) carp (Cyprinus carpio L.) × goldfish (Carassius auratus L.) hybrids and characteristics of their offspring

The purpose of this study was to investigate reproductive ability of backcross triploid koi (Cyprinus carpio L.) × goldfish (Carassius auratus L.) hybrids. These triploids have been obtained by crossing of F₁ hybrid females producing diploid eggs with males of parental species. Triploid hybrid females, when crossed with goldfish or koi males, produced mostly aneuploid fish with ploidy range from approximately 2.2n–3.2n with a mean value 2.5n; some fish in crosses of triploid females with koi males were tetraploid (4.0n). Since analysed fish had in their genomes one haploid set from parental males, the data indicate that triploid hybrid females mostly produced aneuploid eggs with ploidy range from approximately 1.2n–2.2n and a mean ploidy around 1.5n while some eggs were triploid. Triploid hybrid males were completely sterile and have not released any sperm after hormonal injection. Despite their low viability, some aneuploid fish obtained from triploid hybrid females were raised in indoor recirculating systems until the age of 2 years and their reproductive ability has been evaluated. One aneuploid female with ploidy 2.1n produced larvae with ploidy range from 2.9n to 3.4n with a mean ploidy of 3.1n when crossed with a koi male; about 60% of obtained larvae had ploidy from 3.0n to 3.2n. These data indicate that this female produced mostly eggs with unreduced ploidy level.     

Studies on sperm of diploid and triploid tench, Tinca tinca (L.)

The tench Tinca tinca is an interesting fish from the viewpoint of polyploidy and related atypical reproduction aspects. Triploid tench were produced artificially. Studies of spermiation as well as of sperm motility and structure were performed on several triploid and diploid males simultaneously with individual experimental crosses with diploid females to define their reproductive capacities. The testes of triploids visually looked less developed in the most of cases with lower sperm production (0.05 cm³ sperm per male), GSI and weight of testes compared to diploids (0.58 cm³ sperm per male). Analysis of variance showed significant influence of ploidy level on the percentage of motile spermatozoa. Triploidy did not change percentage of live spermatozoa and velocity of spermatozoa at the first time of sperm movement. The study of sperm structure by scanning electron microscopy revealed that most sperm cells were of normal structure with some anomalies. Sperm heads of triploid and diploid males were mostly round-shaped, 1.86±0.2 and 1.6±0.18 μm in diameter. The midpiece of triploid spermatozoa was slightly narrower than that of diploid ones with typical cylindrical shape. Flow cytometry revealed sperm cells of triploids to be largely aneuploid (1.47 n) with high mosaic DNA, oscillating from haploid DNA content (1.0 n) to diploid DNA content (1.9 n). Experimental crosses between triploid males and diploid females revealed that these males were capable to stimulate effective development with relatively high level of fertilization and hatching rates from 0 to 70%. In conclusion, triploidization does not seem to guarantee sterility of tench.     

Comparative performance of growth,vertebral structure and muscle composition in diploid and triploid Paralichthys olivaceus

Growth, skeletal structure and muscle composition of cold‐shock‐induced triploid olive flounder Paralichthys olivaceus were investigated. The average values of total length and total weight of triploids were higher than those of diploids from 5 to 11 months posthatch (mph). The growth difference disappeared after 11 mph. The skeletal structure of flounder at 11 mph was observed by X‐ray imaging method. There are four kinds of vertebral deformity including vertebrae fusion, one‐sided compression, two‐sided compression and vertically shifted. The trunk region (V8–18) and tailing end of the vertebral column were the predominant locations of deformity. In general, the frequencies of vertebral deformities in triploids (60.0%) were higher than those in diploids (33.3%, p < 0.05). Both the number of fish with deformed vertebrae and the average frequencies of deformed vertebrae in triploids were significantly higher than those in diploids (p < 0.05). The muscle tissues of diploid and triploid flounder at 11 mph contain the same types of fatty acid and free amino acid profiles. The number of fatty acids with significant higher contents in diploids and triploids was one and ten, respectively (p < 0.05). The contents of free amino acids showed no difference between triploid and diploid fish.     

The reproductive physiology of three age classes of adult female diploid and triploid brook trout (Salvelinus fontinalis)

Triploid female fish show impaired gametogenesis and are unable to produce viable offspring. The reproductive physiology of artificially-induced triploid female salmonids has been well described up until the time of first sexual maturation in diploids, but few reports exist for older triploids. This study reports the influence of triploidy on growth, ovarian development and reproductive endocrinology among three age classes of female brook trout (Salvelinus fontinalis) in comparison to sibling diploids. Triploids were larger than diploids for most of the study period, but the difference was statistically significant only during maturation and spawning of 2⁺ diploids. Plasma estradiol-17 (E₂), testosterone (T) and vitellogenin (VTG) levels in triploids were generally lower than in diploids, and VTG was the only parameter to show seasonal fluctuations resembling those of diploids. Triploids showed significantly lower GSI and total oocyte number than diploids of similar age, and only half of all triploids sacrificed during the study (n=56) had developing oocytes in their ovaries. At age 3⁺, 13 of 19 triploid females had oocytes at various stages of development, including perinucleolar, yolk vesicle and yolk globule stages. In addition, three of these fish had collectively produced 72 mature stage oocytes. Thus, whereas diploid brook trout can produce mature oocytes as two-year-olds, triploids cannot do so until four years of age, with the number of mature oocytes being greatly reduced.     

Effect of diet on the lipid composition of the commercial clam Donax trunculus (Mollusca: bivalvia): sex‐related differences

This paper describes the lipid composition of the commercial bivalve Donax trunculus and the differences originated when the animals were fed with two phytoplankton species (Tetraselmis suecica and Chaetoceros sp.) in a hatchery. We also analysed sex‐related differences in lipid classes and fatty acid profile. Total lipids were higher in females than in males. Triglycerides and phospholipids were the major lipid components, and the former dominated in females and the latter in males. The main fatty acids in both sexes were 16:0, 20:5n‐3 (eicosapentaenoic acid) and 22:6n‐3 (docosahexaenoic acid). Females showed higher percentages of saturated and monounsaturated fatty acids and lower levels of polyunsaturated fatty acids than males. Significant differences in total lipid, lipid classes and fatty acid profiles were also found due to diet. Docosahexaenoic acid decreased and total lipids, free fatty acids, arachidonic acid and EPA increased in both sexes. Despite these differences, the condition of the species was maintained and the broodstock even maturated.     

Seasonal Variations in Fatty Acids of Striped Bass Morone saxatilis

The polyunsaturated fatty acids of the n-3 family associated with fish and fish oils have been shown to have beneficial effects on prevention of heart disease in humans. However, in considering the potential health benefits of fish oil in human health, it is important to realize that the fatty acid composition and total lipid content of fish vary greatly. These variations may be due to species differences or variations within a species due to temperature, diet, developmental stage, sex or seasonal differences. The purpose of the study described here was to determine the effects of season, size class, and sex on the Occurrence of individual fatty acids in striped bass (Morone saxrrtilis). Fish were collected in June, July, August, September, and October. Filets with skin were analyzed for total lipid content. Methyl esters of fatty acids were separated and identified using capillary gas chromatography. Total mean lipid varied from as low as 12% to as high as 30%. There were no significant changes in total crude lipid content due to month collected or sex. However, total lipid content of filets increased significantly (P ≤ 0.05) as size increased. The fatty acids 18:1n–9 and 16:0 made up the majority (62%) of the fatty acids present in any season, size class, or sex. The monounsaturated fatty acids dominated in all fish. There were no significant differences in individual fatty acids due to sex. However, fish size had a significant influence on fatty acid composition. Overall, saturated fatty acids decreased as size increased, while polyunsaturated fatty acids increased or remained constant. There were no significant changes in mean percent saturated or monounsaturated fatty acids with respect to month when controlling for size. However, total percent polyunsaturated fatty acids decreased significantly in October compared to August and September. This overall decrease was due mainly to a decrease in 22:6n–3. Conversely there was a significant increase in 16:11n–9 in October. All other fatty acids remained constant. When data from this study were compared to fatty acid data from the literature, striped bass was not as good a source of 20:5n–3 and 22:61n–3 as sardines, anchovies or mackerel.     

The growth and reproductive endocrinology of adult triploid Pacific salmonids

This paper describes the effect of triploidy on growth and reproductive endocrinology in the months leading up to and including spawning in rainbow trout,Salmo gairdneri, and pink salmon,Oncorhynchus gorbuscha. Growth rates were the same for diploid and triploid rainbow trout, but triploid female pink salmon were smaller than maturing diploid females and diploid and triploid males of the same age. Triploid males of both species developed typical secondary sexual characteristics and had normal endocrine profiles, although their cycle appeared to be delayed by about one month. Triploid females remained silvery in appearance and showed no endocrine signs of maturation, even at the level of the pituitary. Thus, although triploids of both sexes are genetically sterile, only the females do not undergo physiological maturation.Reported in part at the Third International Symposium on Reproductive Physiology of Fish, St. John's, Newfoundland, August 2–7, 1987.     

Growth and reproductive performance of triploid and diploid blacklip abalone, Haliotis rubra (Leach, 1814)

Growth and reproduction of triploid and diploid blacklip abalone Haliotis rubra (Leach, 1814) were compared in a 30-month study. Triploidy was induced by inhibition of the second polar body formation using 6-dimethylaminopurine (6-DMAP) or cytochalasin B (CB). There were no significant differences in growth and survivorship between triploid and diploid abalone. However, triploid abalone had a more elongated shell and greater foot muscles than diploid abalone. A slightly curvilinear growth in shell length was conformed to all treatments. While diploid abalone had reached sexual maturity and spawned during the study, gonadal development and gamete maturation were abnormal in triploids. Female triploids lacked an apparent gonad at the macroscopic level but microscopic examination revealed that they had a thin layer of oogonia development. In contrast, male triploids were able to form similar-sized gonads to diploids during most of the reproductive period, but with brown-yellow discolouration and stalled gametogenesis at spermatocyte formation. Sex ratio of triploid abalone did not deviate from 1:1. With the onset of sexual maturation, growth and gonadal maturation occurred concurrently in diploid abalone, and there was no indication that growth of (diploid) abalone was reduced.     

Alterations in enzyme activities in vital organs of triploid female catfish Heteropneustes fossilis (Bloch)

Triploid Heteropneustes fossilis (Bloch) showed sterility and higher growth potential than the normal diploid fish. Activities of some metabolic enzymes such as cytosolic NADP-malate dehydrogenase (NADP-MDH), mitochondrial NAD-malate dehydrogenase (NAD-MDH) and glutamate pyruvate transaminase (GPT) were evaluated in liver, brain and kidney along with glucose-6-phosphate dehydrogenase (G-6-P D) in ovary of female triploid catfish. Activities of these enzymes showed distinct seasonal periodicity, mostly with highest activities in prespawning and spawning periods, in both diploid and triploid catfish but differed in magnitude. In triploid liver, GPT showed higher activity than the diploid counterpart in prespawning and spawning periods. On the contrary, mitochondrial NAD-MDH and cytosolic NADP-MDH in this organ showed a consistent lower activity than the diploid in all stages or in some stages of reproductive cycle respectively. Interestingly, none of the enzymes in brain and kidney of triploid female catfish showed significant changes in comparison to the diploid counterpart. The triploid ovary maintained a significantly lower level of G-6-P D activity throughout the resting, preparatory and pre-spawning periods compared to the diploid ovary. Lower level of malic enzymes (NAD-MDH and NADP-MDH) in liver and G-6-P D in ovary are in close synchrony with lower level of estradiol-17β in plasma of female triploids as found in earlier study.     

The fatty acid compositions of total,neutral and polar lipids in wild and farmed lambari (Astyanax altiparanae) (Garutti & Britski, 2000) broodstock

One way to develop broodstock fish diets is to determine the compositions of wild broodstock tissues and attempt to replicate these compositions in the eggs of farmed fish via dietary manipulation. We collected 30 wild and 30 farmed lambari females that were in the reproductive stage. The extraction and separation of polar and neutral fractions and the saponification and methylation of lipids were performed and analysed in the muscles, livers and ovaries to determine the compositions of the fatty acids via gas chromatography. Regardless of habitat, lambaris mobilize large amounts of fat to the ovaries during the reproductive period, in addition to highly unsaturated fatty acids, such as arachidonic, eicosapentaenoic and docosahexaenoic acids. The wild lambaris were observed to contain higher levels of various fatty acids, including linoleic acid, which is an essential fatty acid. The most abundant fatty acid that was observed in the commercial diet was linoleic acid, which was supplied in all fish farmed tissues. The commercial diet has low AA, EPA and DHA contents, and, higher levels of these fatty acids were recorded in the tissues of farmed lambari, which suggests that this species are able to elongate and desaturate precursors, linoleic and linolenic acids, into highly unsaturated fatty acids.     

Induction of triploidy in mud loach (Misgurnus mizolepis) and its effect on gonad development and growth

Triploidy was induced in mud loach (Misgurnus mizolepis) by cold shocking fertilized eggs 5 min post-fertilization at 2°C for 15 to 60 min. Best results were obtained when eggs were shocked for 60 min; 98% of fish examined in that treatment were triploids. Triploidy was confirmed by erythrocyte measurements and chromosome counts. Diploids had 48 chromosomes, while triploids had 72. Histological analysis of 9-month-old triploid ovaries showed an appreciable number of oocytes at the chromatin nucleolus stage with considerable interstitial tissue. However, diploids had well developed oocytes. Diploid testes from diploid males exhibited normal spermatids and spermatozoa, while a few were seen in triploid males. Growth rate was evaluated over a 9-month growth trial. Although male and female triploids were slightly heavier than their diploid counterparts from the third to the ninth month, their growth rates were not significantly different compared to their diploid controls.     

The Effect of Thermal Treatment Method on Fatty Acid Composition in Northern Pike (Esox lucius) Fillets

The aim of the present study was to investigate the effect of thermal treatment methods, such as microwave cooking, steaming, and shallow pan-frying on fatty acids composition in northern pike fillets. All methods of thermal treatment increased fat content in the fillets. Microwave cooking decreased the proportion of saturated fatty acids and increased the proportion of total, n-3, and n-6 polyunsaturated fatty acids as compared with fresh fillets. Steam cooking did not affect the proportion of saturated, monounsaturated, and polyunsaturated fatty acids, whereas pan-frying significantly increased the proportion of monounsaturated and n-6 polyunsaturated fatty acids. All cooked fillets, regardless of the thermal treatment used, had acceptable n–6/n–3 ratio; however, microwave cooking enabled fish products to obtain more beneficial n-6/n-3 ratio than frying. Due to high polyunsaturated fatty acids content and low n-6/n-3 ratio, microwave cooking may be recommended for preparing wild pike fillets in households.     

Reproductive performance and mature gonad morphology of triploid and diploid Black Tiger shrimp (Penaeus monodon) siblings

Sibling harvest age Black Tiger shrimp triploids and diploids of both sexes were reared to reproductive maturity, crossed with wild caught females and males, conditioned for spawning and a comprehensive reproductive performance trial was undertaken. Ovarian development, spawning frequency, fecundity, hatch rate, gonad morphology, male reproductive tracts and thelycum impregnation rates of the wild female × triploid male cross were assessed. After ablation, ovarian development and cycling between wild G₀ diploid and G₁ diploids was not significantly different, whereas G₁ triploids failed to show any signs of ovarian development and cycling, thus resulting in no G₁ triploid female spawnings. There were 10 G₀ diploid female × G₀ diploid male first‐spawnings and 9 G₀ diploid female × G₁ diploid male first‐spawnings, all of which produced viable nauplii. In comparison, there were 7 G₀ diploid female × G₁ triploid male first‐spawnings, none of which produced viable nauplii. The 26 wild G₀ diploid female spawnings had more eggs than the 1 G₁ diploid female spawning. Gonad morphology and male reproductive tract assessments showed impaired reproductive development in triploid gonadal tissues of both sexes (compared with sibling diploids and wild shrimp) to a point where complete maturation had not occurred. The thelycum of 16 wild G₀ diploid females crossed with G₁ triploid males had no visible spermatophore present, suggesting that G₁ triploid males are incapable of developing viable spermatophores and mating with females. This study demonstrates that the triploid females and males are incapable of producing viable gametes and are thus reproductively sterile.     

Aurantiochytrium sp. meal as DHA source in Nile tilapia diet,part II: Body fatty acid retention and muscle fatty acid profile

Nile tilapia juveniles (8.35 ± 0.80 g) were fed on four levels (0.0%; 0.5%; 1.0%; 2.0%, 4.0%) of Aurantiochytrium sp. meal (ALL‐G‐RICH?), a source of docosahexaenoic acid (DHA). The 1% Aurantiochytrium sp. meal diet was compared to a control diet, which contained the same amount of DHA as cod liver oil (CLO) at 1.7% diet. Groups of 25 fish were stocked in 100 L tanks and fed twice daily until apparent satiation, for 57 days, at 28°C. Increasing dietary Aurantiochytrium sp. meal reduced the body retention of DHA and n‐3 polyunsaturated fatty acids (n‐3 PUFA) but increased the body retention of alpha‐linolenic (α‐LNA), linoleic (LOA) and n‐6 polyunsaturated fatty acids (n‐6 PUFA). Fatty acid profile in tilapia muscle was affected by increasing dietary inclusions of Aurantiochytrium sp. meal, with an increase in DHA, α‐LNA, n‐3 PUFA and n‐3 long chain‐polyunsaturated fatty acids (n‐3 LC‐PUFA) but a decrease in monounsaturated fatty acids (MUFA), n‐6 PUFA and n‐6 long‐chain polyunsaturated fatty acids (n‐6 LC‐PUFA). There was a larger body retention of DHA, α‐LNA, LOA, n‐3 PUFA and n‐6 PUFA fatty acids and a higher percentage of DHA, n‐3 PUFA and n‐3 LC‐PUFA in muscle fatty acid profile in fish fed on CLO diets than in those fed on 1% Aurantiochytrium sp. Therefore, Aurantiochytrium sp. meal is an alternative source of DHA for Nile tilapia diets.     

Variations in fatty acids composition of pikeperch (Sander lucioperca) liver with respect to gonad maturation

Abstract –  We examined seasonal variations in liver condition factor, total lipid and fatty acid composition of maturing pikeperch ( Sander lucioperca) from Lake Eğirdir, Turkey. The spawning of pikeperch in Eğirdir Lake started in March, continued very fast during May and gradually ended in July as determined by measuring the gonadosomatic index. The total lipid content of male and female liver reached its highest level in September and November (the end of dense feeding period during which temperature decreased sharply), but declined to the lowest level in May (just after spawning). The highest level of liver condition factor was observed in January and March during which gonads mature rapidly. Palmitic acid was the predominant saturated fatty acid (SFA) in female and male liver. The ratio of monounsaturated fatty acids (MUFA) was higher than that of polyunsaturated fatty acids (PUFA) in both sexes. Docosahexaenoic acid (22:6 n -3), eicosopentaenoic acid (20:5 n -3) and arachidonic acid (20:4 n -6) were the predominant PUFA. Seasonal variations in PUFA were more evident than that in other fatty acids. PUFA, especially the n -3 fatty acids ratio, increased to the maximum level during the sharp decreases in temperature (in November). However, the amount of PUFA decreased to the lowest level in reproductive period. The results suggest that the ratio of pikeperch liver fatty acids is influenced by gonad maturation and temperature variations and that pikeperch requires a large amount of PUFA and the n -3 fatty acids for the development of gonads.     

Novel Meals from Eastern Canada’s Marine Fishery By-products as Alternatives to Fishmeal

There is a current trend in the aquaculture industry to find “eco-friendly” alternatives to fishmeal that will provide fish with all the needed essential nutrients. Our objective was to identify, produce, and chemically characterize nine novel meals composed of underutilized marine fishery by-products from eastern Canada. Snow crab hepatopancreas was found to be a good supplement of monounsaturated fatty acids, iron, copper, and zinc. Herring immature ovaries and shrimp mince could be a partial replacement for fishmeal as revealed by their essential amino acid profile and polyunsaturated fatty acids. Finally, herring testis was very rich in high arginine and taurine.     

Reproductive ability of second generation ornamental (koi) carp (Cyprinus carpio L.) × goldfish (Carassius auratus L.) hybrids and characteristics of their offspring

The purpose of this study was to investigate the reproductive ability of second generation (F₂) koi (Cyprinus carpio L.) × goldfish (Carassius auratus L.) hybrids. Only diploid F₂ males and females were fertile and used in crosses. A significant increase was recorded in male fertility in F₂ versus F₁. In contrast with an earlier study in which only one fertile F₁ male was found, about 20% of F₂ males produced sperm. The observed reproductive ability of F₂ hybrids was similar to that demonstrated by the only fertile F₁ male and F₁ females. F₂ males produced diploid spermatozoa and generated triploids when crossed with koi females. All triploid fish in these progenies were males indicating that F₂ males had a sex chromosome constitution of XY. F₂ females produced diploid eggs and generated mostly triploids when crossed with koi males. In progenies obtained by crosses of F₂ males with F₁ and F₂ females, most of the surviving juveniles (63%–100%) were diploid; a minority of juveniles were aneuploid (ploidy ranged from 2.1n to 3.6n). Diploid fish in these progenies were presumably the result of spontaneous androgenesis and gynogenesis, by the same mechanisms observed earlier in progenies obtained by crossing the F₁ male with F₁ females.