Effect of triploidization on juvenile African catfish (<Emphasis Type="Italic">Clarias gariepinus</Emphasis>)

Triploid African catfish (Clarias gariepinus) produced through cold shock 3 min post fertilization were compared to diploid offspring of the same parentage at 66 days of age. Triploid fish were significantly shorter (11%) and weighed less (18%) than diploids but showed no significant difference in mortality or cannibalism, which can be an important source of losses under aquaculture conditions. Erythrocytes of triploid fish were more oval shaped than the normal spherical shape of diploid erythrocytes, were significantly larger (38%) and had nuclei that were significantly larger (25%) than observed in diploid fish. Erythrocyte morphological characteristics show potential as an indicator for the detection of triploidy in African catfish.     

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.     

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.     

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…     

Characterization of triploid hybrid groupers from interspecies hybridization (Epinephelus coioides ♀ × Epinephelus lanceolatus ♂)

Interspecies hybridization is widely used in aquaculture as a beneficial strategy. Diploid and triploid hybrids have been detected from the interspecies hybridization of Epinephelus coioides ♀ × Epinephelus lanceolatus ♂. This is the first report of triploidization through hybridization in grouper. Confirmation has been obtained through flow cytometry, karyotyping and erythrocyte nuclei measurement. The chromosome numbers of E. coioides, E. lanceolatus, diploid hybrid grouper are 48 and triploid hybrid grouper are 72. Measurements of erythrocyte nuclei indicate that triploid fish have a larger nuclear surface than the diploid groupers, and the average ratio of triploid to diploid surface area is 1.59. During the first 1.5 years, triploid hybrid groupers grow faster than diploid hybrid groupers or either parent species. The average growth rate of triploid hybrids is 1.61 times greater than that of diploid hybrids at 6 months of age and 1.43 times greater at 18 months of age. The triploid hybrid groupers are inferior in gonadal development, with no primary‐growth‐stage oocytes appearing in the gonads at 18 months of age. Morphological studies indicate that triploid hybrid groupers have distinctive differences in snout length, eye diameter, body trunk shape, and tail shape development compared with diploid hybrid groupers. Triploid hybrid groupers have an advantage in growth ability, and artificial breeding of triploid groupers might be of great potential use in the grouper aquaculture industry.     

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.     

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.     

Effects of the estrogen mimic genistein as a dietary component on sex differentiation and ethoxyresorufin-O-deethylase (EROD) activity in channel catfish (<Emphasis Type="Italic">Ictalurus punctatus</Emphasis>)

A number of aquaculture species, including channel catfish Ictalurus punctatus, are fed high proportions of soybean meal in their diet. We have investigated the potential for the most common phytoestrogen in soybean meal to alter phenotypic sex during sexual differentiation in channel catfish. Channel catfish were fed four dietary concentrations of the phytoestrogen genistein (0, 2, 4, and 8 mg g⁻¹) to determine its effect on gonadal sex differentiation. The four treatment diets were fed to sexually undifferentiated channel catfish between 5 and 140 days post hatch (dph) and between 60 and 150 dph. Phenotypic sex was determined by histological examination of the gonads. Ethoxyresorufin-O-deethylase activity was not significantly different among the treatment and control groups. Phenotypic sex was significantly dependant on dietary phytoestrogen concentration (P = 0.01). Additionally, logistic regression showed a significant relation between genistein concentration in the diet and gonadal sex (P = 0.02). Intersex individuals were present at all treatment concentrations, with increasing proportions of intersex fish as the genistein concentration increased for individuals fed treated diets between 5 and 140 dph. Increased proportions of phenotypically male individuals resulted from chronic dietary exposure to the estrogen mimic genistein. There were no significant differences in the proportions of males and females between feed treatment durations. These findings establish that dietary concentrations of genistein can alter sex ratios in cultured channel catfish populations and demonstrates the need to further understand the actions of this and other prominent phytoestrogens in aquaculture species.     

Cryopreservation of Paddlefish Polyodon spathula Milt

Abstract.— A practical procedure for cryopreserving milt of paddlefish Polyodon sparhula was developed to obtain thawed spermatozoa that would fertilize eggs and permit hatching of normal larvae. Milt was mixed with a cryoprotectant medium containing DMSO (2.4 M) in a ratio of 3:1(milt: medium; final concentration of DMSO 0.6 M), stored in 5.0-mL freezing straws, and frozen in dry ice (15 min) and then in liquid nitrogen. A total of three replicates were made; the milt of a different male was used in each replicate. Motility of the thawed spermatozoa decreased to 50%-25% as compared to 100% motility of the fresh (control) spermatozoa. Hatching of paddlefish (16.3 ± 2.2%) from eggs fertilized with thawed spermatozoa was significantly lower ( P ≤ 0.01) than the hatch rate (90.8 ± 2.5%) for the control. It was suggested that an increase in viable motile spermatozoa to egg would result in better fertilization and hatching of paddlefish.     

A comparative study of growth and feed utilization efficiency of sex-reversed diploid and triploid Nile tilapia, Oreochromis niloticus L.

Growth performance, survival and feed utilization of diploid (2n) and triploid (3n) sex‐reversed male and female Nile tilapia were evaluated at maintenance feeding (1% body weight (BW) day^?1), fixed feeding (3% BW day^?1) and apparent satiation feeding levels in a freshwater recirculation system comprised of thirty‐six 1‐m³ concrete tanks at the Asian Institute of Technology, Bangkok, Thailand. Triploid Nile tilapia (3n) was produced by subjecting fertilized diploid (2n) tilapia eggs to heat shock. After hatching, fish were sex‐reversed to all‐male and all‐female populations by oral administration of 17 α‐methyltestosterone (60 mg kg^?1 feed) and ethynylestradiol (100 mg kg^?1 feed) respectively. There was significantly higher growth with increased ration levels in both male and female groups. There were no significant differences in final BW, specific growth rate, survival rate, feed conversion ratio and protein efficiency ratio between diploid and triploid fish. Triploids had lower gonad weights than diploids, and this was particularly evident at the satiation feeding level. Triploid fish had a significantly higher apparent net protein utilization and percentage of gutted weight than diploids at all feeding levels. Higher protein utilization efficiency of triploids might be an advantage for commercial tilapia culture but further research is necessary to make such a conclusion.     

The Influence of Ovarian Fluid pH of Stripped Unfertilized Channel Catfish,Ictalurus punctatus,Eggs on the Hatching Success of Channel Catfish ♀ x Blue Catfish,Ictalurus furcatus♂, Hybrid Catfish Eggs

This study was designed to determine the effect of ovarian fluid pH of stripped unfertilized channel catfish, Ictalurus punctatus, eggs on fertilization and hatch rate of channel catfish ♀ x blue catfish, Ictalurus furcatus♂, hybrid catfish eggs. A significant correlation was established between ovarian fluid pH of stripped channel catfish eggs and hybrid embryo hatch rate (R² = 0.75, P = 0.01), suggesting ovarian fluid pH of stripped catfish eggs prior to fertilization can be predictive of the hatching success of hybrid catfish embryos. These data were used to categorize pH of stripped eggs: pH <7.0 as “low pH eggs,” pH 7.0–7.4 as “medium pH eggs,” or pH >7.4 as “high pH eggs” quality eggs. The range in percent hatch rate for these pH categories was <15%, 15–30%, and >30%, respectively. Higher calcium concentrations during incubation do not appear to improve hatching success of “low pH eggs.” The predictive model presented herein describes a quick method for evaluating stripped catfish eggs for hybrid fry production in catfish hatcheries.     

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.     

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.     

Changes in free amino acid profile of red snapper <Emphasis Type="Italic">Lutjanus campechanus</Emphasis>, eggs,and developing larvae

The free amino acids (FAA) profile was determined for newly fertilized eggs and resultant larvae from wild-caught red snapper Lutjanus campechanus induced to spawn with hCG. Yolk sac and oil globule volumes of eggs and larvae were monitored over time from digital photographs. FAA profiles of the eggs and larvae were measured in picomoles (pmol) of FAA/mg of eggs by HPLC. Newly fertilized eggs had a mean total FAA content of 21.72 ± 3.55 nmoles/egg (92.81 ± 9.71 nmoles/mg eggs). Leucine, valine, lysine, and isoleucine were the most abundant essential FAA comprising 35.9% of the total FAA. Alanine, serine, asparagine, and glycine were the most abundant non-essential FAA comprising 34.2% of the total FAA. At 24 h post-hatch (hph) the mean total FAA had decreased by 81% since egg fertilization. The bulk of the FAA decrease was between the time of hatch and 12 hph. Only 8.5 ± 1.5% of the initial concentration in fertilized eggs of isoleucine, 9.7 ± 2.5% of arginine, and 9.9 ± 2.0% of threonine remained at 12 hph. Among the non-essential FAA, alanine dropped the most by 12 hph with 4.6% of the concentration found in a recently fertilized egg remaining, while cysteine had increased 254.7 ± 26.2%. The yolk sac volume decreased rapidly in the first 12 hph and was further reduced 77.0 ± 2.5% from 12 to 24 hph. The oil globule depletion rate was a more linear decline from fertilized egg to 36 hph.     

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.     

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.     

A comparative study of the survival,growth and gonad development of the diploid and triploid Hong Kong oyster,Crassostrea hongkongensis (Lam & Morton 2003)

The Hong Kong oyster, Crassostrea hongkongensis is an economically important and widely cultivated oyster species in southern China. Aiming to improve the aquaculture potentials of C. hongkongensis, a triploid organism was generated by inhibiting the second polar body formation in the fertilized eggs, using 0.5 mg L^?1 cytochalasin B. Triploids exhibited significantly higher growth and were larger than the diploids throughout the life cycle. Much of the triploid growth advantage can be attributed to resource reallocation to structural development due to gonadal sterility. Most of the triploid oysters exhibited polymorphic sterility with atrophic gonads and abnormal gametogenesis. Only few ‘dysfunctional’ sperms were produced by the triploid males, thus ruling out reproductive and genetic contamination of natural oyster populations from triploid cultivations. This advantage in addition to the superior growth of triploids makes it a potent alternative in Hong Kong oyster aquaculture industry. Triploid survival was mostly found to be similar to that of diploids except for day 360, when a significant triploid advantage in terms of survival was noted. Furthermore, we found that growth and survival of Hong Kong oyster is significantly higher at the Zhuhai site in comparison to the Beihai site. Our results indicate a strong environmental effect in Hong Kong oyster cultivation and points to the fact that Zhuhai site was more suitable for oyster farming than the Beihai site. Overall, this study provides valuable information to the aquaculture industry of southern China.     

三倍体鲶鱼的形态学性状及生长

采用静水压休克法人工诱导鲶鱼受精卵，使三倍体出现率达100％，其主要形态学性状同二倍体鲶鱼比较研究表明，三倍体鲶鱼具有体形大，相对体高及丰满度均较高，体长差异不明显，外部形态及摄食仍保留着二倍体鲶鱼的特征，但比二倍体鲶鱼增长快，在池塘养殖90天后，体重比二倍体鲶鱼平均增加39．71％。试验证明三倍体鲶鱼是具有养殖生产前景和推广价值的新对象。