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.     

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.     

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.     

Preliminary study on performance of triploid Thai silver barb, Puntius gonionotus

The triploid chromosome condition was induced in Thai silver barb (Puntius gonionotus) by application of cold shock (2°C) to eggs at time intervals after activation of 0.5 min with a duration of 10 min which resulted in mean triploidy yield of 72.5% at 9 months of age. Growth rate of the 2–9-month-old, cold shock group (0.1–6.2 g/month) did not differ from that of the control (0.1–5.7 g/month). Gonadal somatic indices (GSI) of presumed triploid males and females were lower than that of control (GSI values of the presumed triploids were 35.0–60.2% and 28.7–75.9% of control males and females, respectively). Spermatogenesis and oogenesis were retarded in triploids. However, all stages of spermatogenic cells were observed in triploid males, including few spermatozoa. Oocytes of triploid females did not undergo vitellogenesis while normal oogenesis was observed in diploids. Nuclear volume of red blood cells (RBCs) of triploid fish was 1.63 times larger than that of diploids.     

Triploid Induction in the Yellowtail Tetra,Astyanax altiparanae,Using Temperature Shock: Tools for Conservation and Aquaculture

Triploidization is an interesting tool to produce sterile fish. In the yellowtail tetra, Astyanax altiparanae, this can be applied for aquaculture and surrogate technologies. In this study, we compared the efficacy of cold (2 C) or heat shock (38 C, 40 C, and 42 C) on triploid induction in the yellowtail tetra. The eggs were treated with cold or heat shock, 2 min postfertilization (30 min in cold shock or 2 min in heat shock). Intact embryos served as the control group. Ploidy status was confirmed by karyotyping, flow cytometry, and nuclear diameter of erythrocytes. The hatching rate decreased after cold shock (12.69 ± 15.76%) and heat shock at 42 C (0.35 ± 0.69%) in comparison with the control group (63.19 ± 16.82%). At 38 C and 40 C, hatching rates (61.29 ± 17.73% and 61.75 ± 22.1%, respectively) were not decreased. Only one triploid arose at 38 C (1/80). At 40 C, a high number of triploids arose (72/78). At 42 C, very few embryos developed into the hatching stage. A large number of haploid individuals arose after cold shock (61/75), with only one triploid. Our results indicate that heat shocking of embryos at 40 C is optimum for triploid production in the yellowtail tetra.     

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.     

Growth and Survival of Intrastrain and Interstrain Rainbow Trout (Oncorhynchus mykiss) Triploids1

Triploid rainbow trout exhibit improved survival and extended growth during sexual maturation, compared to their diploid counterparts. However, there have been few benefits demonstrated prior to sexual maturation. This study was undertaken to investigate the possibility of improving growth and survival parameters in triploids through interstrain crosses. Triploids were induced by heat shocking fertilized eggs from intra- and interstrain crosses of two rainbow trout strains. The four triploid groups and their diploid counterpart groups were reared to 233 days post-hatching and analyzed for growth and survival characteristics. Compared with diploids, triploids had significantly (P < 0.05) higher mortalities during the first 100 days post-fertilization, primarily just prior to and after hatching. However, during the remainder of the study triploids exhibited significantly (P < 0.05) lower mortalities than diploids. During the first 50 days of rearing all four triploid groups were significantly shorter and lighter than their diploid counterparts. The growth of the triploid groups later in the study varied considerably. At the conclusion of the rearing phase, one interstrain triploid group was significantly (P < 0.05) longer than its diploid counterpart, although not significantly heavier. The other triploid groups were either significantly smaller than, or equal to the diploids. Analysis of variance indicated that the growth of triploid rainbow trout was significantly affected by maternal strain effects. These results suggest that the use of specific strains and crosses may improve the growth of triploid rainbow trout.     

The breeding potential and biochemical composition of triploid Manila clams, Tapes philippinarum Adams and Reeve

In 1989 and 1990. triploid Manila clam, Tapes philippinarum Adams and Reeve, seed were reared to 15-20 mm at the Fisheries Laboratory, Conwy, and planted out in the Menai Strait, North Wales. In each of the summers of 1992 and 1993, three of these batches, at 2, 3 or 4 years old, were returned to the laboratory to assess ploidy, size, spawning potential and biochemical composition. Percentage triploidy at this time was similar to that in the seed. After 6 and 8 weeks of warmwater conditioning. Only 45 out of l21 triploids (37%) were induced to spawn by thermal shock, with only one spawning as a male. By comparison, 75% of diploid clams spawned with a 1:1 ratio of males to females. Mean fecundity of triploids was significantly lower than that for diploids, at 0.497 compared with 1.54 million eggs per female. Compared with eggs from diploid females, eggs from triploids were larger and significantly fewer of them developed into D-larvae when fertilized by sperm from diploid males. Triploid clams were heavier and had a higher condition index and carbohydrate content than diploids of the same age, but lipid levels were similar. Potential advantages of producing and cultivating 100% triploid batches of Manila clam seed are discussed.     

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.     

Application of an electrophoretically detectable genetic marker to ploidy testing in brown trout (Salmo trutta L.) triploidised by heat shock

A phosphoglucose isomerase (PGI) variant, of brown trout (Salmo trutta L.), expressed in adipose fin tissue, was incorporated into experimental batches through selective crossing of brood-stock having the PGI-3 (110) marker gene. PGI-3 (110) homozygous females were crossed with PGI-3 (100) homozygous males to yield all PGI-3 (110/100) progeny. These were subject to heat shock for 10 min at 28°C, commencing 10 min after insemination. Electrophoretic examination of fry indicated that diploids and triploids were separable on the basis of PGI-3 banding patterns, having staining ratios of 1:2:1 and 4:4:1, respectively. This confirmed the retention of an extra maternal PGI-3 (110) gene in triploid individuals. Potential applications of this marker in triploid studies in brown trout are discussed.     

Farming triploid oysters

Although the commercial benefits of triploidy have been evaluated in the Pacific oyster Crassostrea gigas (Thunberg, 1793), eastern oyster C. virginica (Gmelin, 1791), Sydney rock oyster Saccostrea glomerata (Gould, 1850) and European flat oyster Ostrea edulis (Linnaeus, 1750), so far this technique has only been commercialised for Pacific oysters.

Commercial production of triploids on the West Coast of North America began in 1985. Since then production of triploids has greatly increased and the use of tetraploid males to fertilise eggs from diploids to produce batches of 100% triploids has been developed. In 1999/2000, triploid Pacific oysters made up 30% of all Pacific oysters farmed on the West Coast of North America. Some hatcheries now use tetraploid males instead of chemical or physical stress to produce triploids. The rapid uptake of triploid and tetraploidy techniques has been facilitated by the almost total dependence that these oyster industries have on hatcheries for the supply of seed. This industry in the Pacific Northwest of the US and in British Columbia, Canada, would not have developed to its current size without hatchery seed supplies. Triploids are preferred over diploids in summer because diploids are less marketable when in spawning condition.

There was only limited interest in triploidy production in France until 1999, when IFREMER began to make sperm from tetraploids available to commercial hatcheries. In 1999/2000, only 10% to 20% of all the hatchery-supplied Pacific oyster spat were triploids, but with the use of sperm from tetraploid oysters, this could increase sharply.

Elsewhere around the world, the commercial uptake of triploid oysters has been slow to develop. However, in countries where the production of Pacific oysters is based on hatchery supply of seed, it is likely that with the use of tetraploid oysters, the farming of triploid oysters will increase in the near future.     

三倍体僧帽牡蛎生殖腺发育观察

在僧帽牡蛎繁殖盛期，详细观察了二倍体，三倍体僧帽牡蛎生殖腺外部形态特征，比较了成熟卵母细胞卵径和核径。结果表明，外观上三倍体生殖腺发育较二倍体差，三倍本成熟卵母细胞卵径和核径分别比地倍体大１９．６％和１７．６％，体积分别比二倍体增加７０．３％和６４．２％，组织切片检查结果，二倍体生殖腺发育正常，三倍体生殖腺发育受阻，大都在增殖期和休止期，一部分个体可发育至生长期和成熟期，并能产生成熟的卵子和精子，     

三倍体大黄鱼的诱导及其对生长、性腺发育的影响

自20世纪50年代以来,采用染色体操作技术人工诱导多倍体在鱼类遗传育种领域已得到广泛的应用。国内外已先后在三棘刺鱼[1]、鲤[2]、水晶彩鲫[3]等30多种鱼类成功获得三倍体。在理论上,由于三倍体性腺发育受阻,其用于性腺发育的能量可全部用于生长。因此鱼类育种学家期望通过诱导三倍体,使经济鱼类生长更快,经济效益更高。但历经30余年的研究,诸家看法仍未统一。一些学者认为三倍体鱼比二倍体生长快[4,5],另一些学者则认为三倍体鱼并不比二倍体生长快[6,7]还有一些学者的研究表明三倍体鱼在性成熟以后比二倍体生长稍快[8,9]。对于三倍体…     

Experimental production of triploid brown trout, Salmo trutta L., using heat shock

Abstract. This study was designed to investigate the potential of heat shock to produce triploidy in brown trout, Salmo trutta L., and to develop a methodof routinely identifying triploids in this species. Triploids were produced in all heat-shocked batches and were identified by the size of their erythrocyte nucleus, which had a volume ratio of 1:1-57 relative to diploid controls. Cytogenetic and flow cytometric analyses confirmed that trout with the larger nuclei were triploid. Heat shock of 28°Cof 10 min duration initiated 5-15 min post-insemination produced high rates of triploidy in experimental batches (88-2-100%), later shocks at 20-25 min producing lower rates (down to 60%). Reproducibilicy of tripioid rates was generally good, a maximum difference between replicates of 21.9% being observed, the majority of differences being considerably less. The highest triploid yield was produced with a heat shock of 28°C for 10 min initiated at 10 or 20 min post-insemination, the difference between replicates being due to variability in survival to hatch. Survival to hatch was generally lower in groups having higher rates of tripioidy.     

Variability in microsatellite DNA markers in gynogenetic and backcross progenies obtained from ornamental (koi) carp (Cyprinus carpio L.) × goldfish (Carassius auratus L.) hybrid females

Inheritance and segregation at five microsatellite loci were studied in diploid gynogenetic and triploid backcross progenies obtained from koi × goldfish hybrid females, which produce diploid eggs. Gynogenetic and backcross progenies were obtained from three individual hybrid females by inseminating eggs with genetically inactivated and intact sperm of parental species respectively; no shock treatments were applied to the early embryos. Complete absence of paternally specific alleles at all investigated microsatellite loci has proven successful genetic inactivation of spermatozoa by irradiation and confirmed gynogenetic origin of progenies. Genotypic segregations at microsatellite loci showed almost complete homogeneity of gynogenetic progenies and their identity to female parents. These results correspond with previous cytogenetic data on the occurrence of premeiotic endomitosis in hybrid females producing diploid eggs. Fish from triploid backcross progenies had genotypes resulting from combination of entire diploid female genome and haploid genome from male.     

Production of triploid eastern little tuna,Euthynnus affinis (Cantor, 1849)

Herein, we developed a triploidization method using spawned eggs collected immediately after spawning of eastern little tuna (ELT), Euthynnus affinis. ELT broodstock induced the spawning by hormonal treatment in the tank, with the resulting spawned eggs being used for the triploidy induction. Under optimal conditions, the mean ± SEM triploidization and hatching rates were 97.2 ± 2.8% and 84.5 ± 10.3%, respectively. Although triploid ELT showed growth performance equivalent to that of diploids, the triploids died at a higher rate than the diploids during 2–4 weeks post‐hatching when triploids and diploids were reared in the same tank. Therefore, we propose that it would be necessary in a practical operation to use triploid‐only ELT seedlings to avoid selective cannibalism by the diploids. The ELT triploids exhibited an all‐female phenotype. Because previous studies have reported that female triploids show a greater probability of sterility than male triploids, this characteristic could be a major advantage. Since this triploidization method, using spawned eggs, can be performed without handling the broodstock, it is possible to avoid the physical damage caused by the process of artificial insemination, making it possible to repeatedly produce triploid populations without valuable broodstock loss. Thus, we have developed an efficient method to produce ELT triploids, although further study is essential to evaluate sterility of the triploid ELT.     

Induction of triploidy in large yellow crocker Pseudosciaena crocea (Richardson, 1846): effects of pressure shocks and growth performance in the first rearing year

The precociously sexual maturation in large yellow crocker Pseudosciaena crocea has become a serious problem. In an attempt to solve this problem, the production of sterile triploids could be an effective strategy. In this study, triploid P. crocea was obtained by subjecting fertilized eggs to pressure shock. Flow‐cytometry analysis was used to assess ploidy level. In terms of triploid rate and hatching rate, the optimal conditions of pressure shock for triploidy induction in P. crocea were 7500 psi for 3 min shock at 3 min after fertilization at 20 °C. With the application of these parameters, 100% triploid fish were produced. During the first rearing year, triploid P. crocea had a similar growth performance compared with its diploid counterpart before the age of 8 months and showed a significant advantage at the age of 10 and 12 months in body weight and body length (P<0.05). At the age of 12 months, the carcass weight of triploids was markedly higher than that of diploid control, and gonadal somatic index was significantly lower than that of their diploid control. During the first rearing year, survival in triploid group was 76.44%, inferior to its diploid control (83.21%).     

Hormonal profile of growing male and female diploids and triploids of the blue tilapia,Oreochromis aureus,reared in intensive culture

Triploidy as a result of thermal shock exposure of fertilized eggs decreases the growth rate ofOreochromis aureus as compared to their diploid controls, but this is due to the higher female ratio present in triploids (86%) and the lower growth rate of females. When females and males are considered separately, the growth rate is not significantly different in diploids and triploids. Since triploidy results in a malfunctioning steroidogenesis in females (mainly testosterone (T) and 17β-estradiol (E₂)), but does not affect the growth rate, it is concluded that female gonadal steroids do not influence growth unless in pharmacological concentrations. These low levels of gonadal steroids are generally accompanied by higher levels of gonadotropin (GtH), but the difference is not always significant. Despite their lower growth rate diploid females have higher plasma concentrations of growth hormone (GH) during several months compared to the triploid females and diploid males. 3,5,3′-triiodo-L-thyronine (T₃) levels, however, are comparable between diploid and triploid females (except for 1 month), but higher in diploid males in 4 of the 5 months studied. 11-ketotestosterone (11kT) is always higher in males. These results indicate that the higher growth rate of males may be related to the high circulating levels of T₃ and 11kT.     

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.     

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.