Differences in metabolism of triploid and diploid Salmo trutta f. lacustris under acclimation conditions and after exposure to stress situations

Differences in metabolism (enzyme activities, metabolites) between diploid and triploid Salmo trutta f. lacustris were investigated under acclimation and stress conditions. Under acclimation conditions enzyme activities differed for 35% of the 27 investigated key regulatory enzymes and temperature optima for 23%. Muscle and liver metabolites related to energy metabolism and diagnostic indices of blood serum were similar, with exception of acetyl‐CoA being increased in triploids. Metabolic rate was lower and gill ventilation rate higher in triploids in comparison with diploids. During the tested stress situations (24 hr endurance swimming, 3 hr exposure to hypoxia in water with 32% oxygen saturation) muscle and liver glycogen decreased and serum and muscle lactate increased in both ploidy levels. Specifically, for triploids muscle adenylate energy charge and phosphocreatine levels decreased after endurance swimming and muscle and liver adenylate energy charge after exposure to hypoxia. Acetyl‐CoA increased in triploids during both stress situations. In summary, there existed differences in metabolism between the two ploidy levels and the energy metabolism of triploids was less balanced under stress.     

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.     

Growth and gut morphology of diploid and triploid juvenile Atlantic cod (Gadus morhua)

The objective of this paper was to compare the growth and gut morphology of juvenile diploid and triploid Atlantic cod (Gadus morhua) reared under similar conditions. Individually tagged 36‐week‐old diploid (mean weight 49.3 ± 13.8 g) and triploid (mean weight 43.6 ± 11.2) juvenile cod were measured at intervals during a 29‐week growth trial. Data for weight, length, condition factor (K), hepato‐somatic index (HSI), gonado‐somatic index (GSI), Relative Gut Length (RGL) and pyloric caeca number were collected and results were analysed in relation to ploidy status, gender and family contribution. At the end of the experiment, only one family (M2xF3) had many representatives with a relatively even distribution of sexes and ploidies. Diploid females were significantly heavier and had higher K than triploid females in the M2xF3 family (body weight 371.2 ± 120.2 vs. 298.4 ± 100.7 g; K 1.1 ± 0.1 vs. 0.93 ± 0.1), but no differences were found between diploid and triploid males. In the other families (pooled data), no differences in body weight were found between the ploidy groups. In general, triploids had a shorter intestine (RGL) and fewer pyloric caeca than their diploid siblings regardless of gender suggesting possible impairments in nutrient utilization and growth.     

Effect of triploidy on muscle cellularity and flesh quality of turbot (Scophthalmus maximus)

Production of triploid turbot is a relevant goal for the aquaculture industry due to the benefits associated with their sterility and growth. In order for a better understanding of the final quality of the species, muscle cellularity and quality parameters of the flesh were studied in seven diploid and seven triploid turbot after sexual maturation. White muscle cellularity was evaluated throughout white muscle cross‐sectional area from the epaxial left muscle section of each fish. To ascertain the flesh quality, several physico‐chemical parameters (moisture, ash, protein, total fat, fatty acids and minerals) were analysed, and textural mechanical properties (springiness, hardness, chewiness, cohesiveness, gumminess and adhesiveness) were determined objectively with a texturometer. Muscle cellularity was different between both groups, such that muscle fibre density was higher for diploid specimens (P < 0.001) and muscle fibres diameters were higher for triploids (P < 0.001). Both ploidy groups showed no significant differences in moisture, ash, protein and total fat content (P > 0.05). All textural properties analysed showed no significant differences between both ploidy groups (P > 0.05). In both ploidy groups, fibre density and fibre diameter showed no significant correlations (P > 0.05) with textural parameters such as springiness, cohesiveness, chewiness and adhesiveness. Ploidy did not affect fatty acid composition, with the exception of eicosapentaenoic acid (C_20:5ω3) which was higher in the triploid group. No significant differences (P > 0.05) existed between diploid and triploid turbot in content of all minerals analysed. According to our results, the ploidy status determines significant variations on muscle cellularity but does not influence the flesh quality of the turbot.     

Effect of triploidy on quality traits of shi drum (Umbrina cirrosa L.) until the second rearing year

Carcass and flesh morphometric, reological and chemical traits of triploid (3n) and diploid (2n) shi drum (Umbrina cirrosa L.) were evaluated through 7 months. Three age groups, 17‐, 21‐ and 24‐month‐old fish, were investigated. The effects of ploidy were statistically evaluated and the weight of fish was included in the model as a covariate because triploids grew less than diploids. As expected, fish weight was found to be significantly correlated with all the investigated morphometric traits, but showed a negative correlation with some chemical (pH) and colour traits (lightness) of raw fillet. In comparison with controls, triploid shi drums were characterized by different morphological traits that involved a slender body shape. In triploids, a reduction in condition factor, backbone weight, dressing index and an increase in the agility index were also recorded. When the commercial size (i.e. over 300 g) was achieved, triploids exhibited larger coelomatic and fillet (dorsal white muscle) fat deposition than diploids. Among reological traits, colour and texture were affected by ploidy; raw fillet lightness and cooked flesh tenderness were higher among triploids in all the investigated age groups. As fish were fed with a restricted feeding regimen, fillet fat deposition was supposed to be limited. Thus, the effects of ploidy on reological traits may be only partially explained by lipid fillet amount and are supposed to be more related to different fibre muscle architecture.     

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.     

Skeletal anomaly assessment in diploid and triploid juvenile Atlantic salmon (Salmo salar L.) and the effect of temperature in freshwater

Triploid Atlantic salmon tend to develop a higher prevalence of skeletal anomalies. This tendency may be exacerbated by an inadequate rearing temperature. Early juvenile all‐female diploid and triploid Atlantic salmon were screened for skeletal anomalies in consecutive experiments to include two size ranges: the first tested the effect of ploidy (0.2–8 g) and the second the effect of ploidy, temperature (14 °C and 18 °C) and their interaction (8–60 g). The first experiment showed that ploidy had no effect on skeletal anomaly prevalence. A high prevalence of opercular shortening was observed (average prevalence in both ploidies 85.8%) and short lower jaws were common (highest prevalence observed 11.3%). In the second experiment, ploidy, but not temperature, affected the prevalence of short lower jaw (diploids > triploids) and lower jaw deformity (triploids > diploids, highest prevalence observed 11.1% triploids and 2.7% diploids) with a trend indicating a possible developmental link between the two jaw anomalies in triploids. A radiological assessment (n = 240 individuals) showed that at both temperatures triploids had a significantly (P < 0.05) lower number of vertebrae and higher prevalence of deformed individuals. These findings (second experiment) suggest ploidy was more influential than temperature in this study.     

Effect of ploidy on the mortality of Crassostrea gigas spat caused by OsHV‐1 in France using unselected and selected OsHV‐1 resistant oysters

The effect of ploidy on the mortality of Crassostrea gigas spat caused by the ostreid herpesvirus (OsHV‐1) genotype μVar was investigated at five sites along the Atlantic coast in France in 2011. Sibling diploids and triploids were produced using either unselected or selected OsHV‐1‐resistant oysters. No significant interactions were found between the factors of environment, genotype and ploidy at the endpoint dates. The mean mortality rates at the sites were 62% and 59% for diploids and triploids, respectively, and the two rates were not significantly different. The mean mortality rates were 33% and 32% for sibling diploids and triploids, respectively, when OsHV‐1‐resistant parents were used and 91% and 85%, respectively, when unselected parents were used. The results were confirmed through other broodstocks tested in 2013. Our study is the first to clearly show that mortality related to OsHV‐1 is similar between diploids and triploids in C. gigas when the same germplasm is used for both ploidy. Furthermore, OsHV‐1 resistance was not substantially altered by triploidization, indicating that the achieved selective breeding of diploid oysters for OsHV‐1 resistance can be translated into improved survival in 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 triploid oysters in Australia: effect of initial size on growth of diploid and triploid Sydney rock oysters, Saccostrea commercialis (Iredale & Roughley)

In a 2-year grow-out trial, triploid Sydney rock oysters, Saccostrea commercialis (Iredale & Roughley), from two initial size grades grew faster (in terms of both mean whole weight and shell height) than the equivalent initial size grades of sibling diploids (P < 0.05). Small size grade triploids caught up with and had significantly heavier (P < 0.05) final whole weights than large size grade diploids after a 2-years grow-out period. The initial size grade had a significant effect on final mean whole weight and shell height for both ploidy types. After the 2-years grow-out trial, the final mean whole weights (but not shell heights) of small and large diploids (35.8 ± 0.6 g and 39.4 ± 0.5 g, respectively) were significantly different (P < 0.05). Small and large triploids grew at a similar rate for the first 18 months despite the significantly (P < 0.05) heavier final mean weight of large grade triploids (48.4 ± 0.8 g and 61.2 ± 0.7 g, respectively). The effect of the initial size grade on subsequent growth of both diploid and triploid oysters which was demonstrated in the present study is of significant commercial value to hatchery and nursery operators as well as growers of single seed oysters. In addition, small-grade triploids appeared to be more valuable in terms of potential growth rate than all diploid grades. There was no significant difference in the final percentage triploidy between small and large grade triploids. A large proportion of diploid/triploid mosaicism was detected in adult oysters.     

Added value from an added chromosome: Potential of producing large fillet fish from autumn to spring with triploid rainbow trout,Oncorhynchus mykiss

Three‐summers‐old all‐female triploid and diploid rainbow trout were compared after one on‐growing season in sea net cages. Slaughter traits of round weight, gutted weight, fillet weight, carcass% and fillet% were measured at three times in November 2017, January and April 2018. The triploid group had lower daily growth coefficient mean (4.25) and higher feed conversion ratio (1.18) than diploids (4.48 and 1.05, respectively) during on‐growing (June–November). In November, no difference of means was found between mature or immature diploids and triploids for any of the weight traits when the effect of vertebrae defects was statistically removed. However, the triploids had attained higher means than mature or immature diploids in gutted and fillet weight by January, suggesting that the loss of muscle mass during early winter was lower in triploids. Sexually maturing diploids (46%) had lower slaughter yield means compared to triploids or immature diploids at each measurement time, and these differences also increased during overwintering. Instead, the means of yield traits remained similar between the triploid and immature diploid groups through the winter. Likewise, fillet redness remained at equally high level in triploids and immature diploids, whereas in maturing diploids this attribute decreased substantially during overwintering. The triploid group had a higher incidence of vertebral defects (12.0%) than diploids (5.3%). The present results demonstrate the potential of triploid trout in producing large‐sized (>2 kg) fillet fish until spring markets. However, more detailed investigations are needed, particularly regarding the animal health and growth efficiency in triploids, relative to their diploid counterparts.     

Growth and gonadal development in diploid and triploid Atlantic cod (Gadus morhua)

Induction of triploidy has been suggested as an effective tool to prevent spawning of farmed fish. This experiment examined the growth potential of triploid cod when reared communally with diploid ones after the juvenile stage. Pressure treatment was used to induce triploidy in a batch of cod eggs in April 2009. The resulting offspring were reared separately from their diploid counterparts until they reached the proper size for PIT tagging. At the age of 8 months, an equal number of 115 diploids (135.5 ± 3.95 g) and triploids (93.6 ± 2.63 g) were communally reared in a circular flow-through tank until the age of 22 months. By the end of this rearing period, diploids (1,002.4 ± 39.9 g) were significantly heavier than triploids (654.6 ± 27.7 g), but the specific growth rate did not differ significantly during the growth trial. Gonadal development at the age of 22 months was also lower among triploids than diploids, especially for females (5.3 and 91.9 %) but also for males (32.5 and 72.7 %). Sterility among female triploids was evident by the reduced size and dysfunctional gonads, but gonadal development in male triploids was less suppressed. Prevalence of body deformities was, however, significantly higher among triploids (62.6 %) than diploids (33.9 %). Higher prevalence of deformities in triploid cod underlines the need for further fine-tuning of the triploidization procedure or finding other methods of sterilization. At present, triploid cod are still far from being established as an alternative for commercial production.     

Triploid hard clams Mercenaria mercenaria produced by inhibiting polar body I or polar body II

Two types of triploid hard clams Mercenaria mercenaria were produced by inhibiting polar body I (PB1) or polar body II (PB2) with cytochalasin B. Treatments were applied at 22–23°C, with PB1 inhibition starting at 4–7 min postfertilization and ending when PB2 was first observed in control groups, and with PB2 inhibition starting at 17–23 min postfertilization and ending when 80% of control eggs released PB2. Triploid induction success was evaluated by chromosome counting in 2–4 cell embryos and by flow cytometry at larval and juvenile stages. PB2 inhibition produced more triploids (82%–100%) than PB1 inhibition (71%–83%), although the difference was not significant (p ≥ .088). Triploid percentages in PB1‐ or PB2‐inhibited groups showed a small but insignificant decline during the first 6 months. At month 3, PB1 and PB2 triploids were not different from their within‐group diploids, but significantly larger than control diploids; PB1 triploids were significantly larger than PB2 triploids (p ≤ .003). At month 6, PB1 triploids were not different from either within‐group or control‐group diploids, while PB2 triploids were significant larger than both within‐group and control diploid; PB1 triploids were smaller than PB2 triploids. At month 16, PB1 and PB2 triploids in one remaining replicate were not different from their within‐group diploids. Overall, this study shows that triploids can be efficiently produced by PB1 or PB2 inhibition, and their growth performance relative to diploids is variable depending on age and replicates or parental genotype.     

Hemoglobin level,metabolic rate,opercular abduction rate and swimming efficiency in female triploid brook trout (Salvelinus fontinalis)

Metabolic (oxygen consumption) rate, opercular abduction rate and tail beat frequency were determined in two strains of diploid and triploid female brook trout (Salvelinus fontinalis) while these fish swam at 0.37±0.02 body lengths per sec in a Blazka respirometer. Total blood hemoglobin level was also measured and opercular condition examined. Total blood hemoglobin levels in diploids and triploids were equal. The opercular abduction rate was the same in diploids and triploids (regardless of whether triploid opercular condition was good or poor) yet triploids had a lower oxygen consumption rate than diploids, indicating that triploids take up less oxygen than diploids per opercular cycle. Tail beat frequency, an indicator of swimming effort, was the same in diploids and triploids, suggesting that triploids require less oxygen than diploids for a similar swimming effort.     

Adult triploid Atlantic salmon (Salmo salar) have higher dietary histidine requirements to prevent cataract development in seawater

The study investigated cataract preventive effects of dietary histidine (His) supplementation in triploid Atlantic salmon during seawater grow‐out. Groups of individually PIT tagged diploid (2N) and triploid (3N) postsmolts were fed one of two supplemented dietary histidine levels; low (L, 12.6 g kg^?1 diet) or high (H, 17.4 g kg^?1 diet) from March to September following their first sea winter. Low severity cataracts were detected in both ploidy prior to supplemented His diet application. Thereafter, 3N‐L showed progression of cataract development in the second spring‐summer period, while development was inhibited in 3N‐H. Severity of cataract showed a strong family effect. A positive correlation between initial triploid seawater growth (weight and TGC) under increasing water temperature and cataract severity was identified as a major risk factor. The relationship was reversed at harvest, where triploids were on average 7.5% smaller than their diploid siblings. Lens N‐acetyl‐histidine content reflected dietary His inclusion level and cataract severity, although no significant differences in lens His content were evident between ploidy or dietary groups. Results indicate that triploid Atlantic salmon appear to have a higher dietary histidine requirement than diploids and that preventative measures can be taken to mitigate further cataract development.     

Effects of temperature on feed intake and plasma chemistry after exhaustive exercise in triploid brown trout (<Emphasis Type="Italic">Salmo trutta</Emphasis> L)

The physiological effect of temperature on feed intake and haematological parameters after exhaustive swimming in diploid and triploid brown trout (Salmo trutta) was investigated. Trout were exposed to an incremental temperature challenge (2 °C/day) from ambient (6 °C) to either 10 or 19 °C. Feed intake profiles did not differ between ploidy at 10 °C; however, triploids had a significantly higher total feed intake at 19 °C. After 24 days, each temperature–ploidy group was exposed to exhaustive swimming for 10 min. The haematological response differed between ploidy, with the magnitude of the response affected by temperature and ploidy. Post-exercise, acid–base and ionic differences were observed. Plasma lactate increased significantly from rest for both temperature and ploidy groups, but glucose increased significantly at higher temperature. Post-exercise, triploids at 19 °C had significantly higher osmolality and cholesterol than diploids, but differences were resumed within 4 h. Elevated alkaline phosphatase (ALP) and aspartate aminotransferase (AST) in fish at higher temperature suggested greater tissue damage; however, both ploidy responded similarly. Despite no significant differences in deformity prevalence, the type and location of deformities observed differed between ploidy (decreased intervertebral space with higher prevalence in tail area and fin regions for diploids, while vertebral compression, fusion in cranial and caudal trunks for triploids). These results suggest triploids have greater appetite than diploids at elevated temperature and that triploids suffer similar blood disturbances after exercise as diploids. These findings have implications for the management of freshwater ecosystems and suggest that stocking triploid brown trout may offer an alternative to diploid brown trout.     

Comparative Growth of Diploid and Triploid Asian Catfish Clarias macrocephalus in Thailand

Wild caught Asian catfish were spawned manually following HCG injection, and a portion of the eggs were subjected to cold-shock at 4 C for 15 min within 2-min post-fertilization. Nuclear diameter measurements of cold-shocked fish revealed that 96% were triploids (3N), while non-shocked fish were all diploids (2N). During larval and fry culture (first 26 d), triploid fish mortality was =50%, while diploid mortality was =25%. Following 8-mo culture in tanks at three stocking densities, triploid fish survival was significantly greater ( P < 0.05), than diploids, with 84.0% and 57.3%, respectively. Triploid live weight was also significantly greater than diploids, with 69.2 and 45.9 g averages, respectively. Ninety-two percent of diploids had welldeveloped gonads after 8 mo; whereas none of the triploids had mature gonads. Gonads were undifferentiated with 31% of the triploids. These sexually undifferentiated fish had greater growth rates than male or female triploids, and greater growth than all diploids. Carcass weight (gutted) of triploids was 95.8% of live weight, compared with 92.5% for diploids. Lastly, triploids had very few deformities compared with diploids, with 1.3% and 17.6%, respectively. Deformities included curved spines, and humped backs just posterior of the head.     

Comparative studies of the growth,survival, and reproduction of diploid and triploid Kumamoto oyster,Crassostrea sikamea

Triploid oysters have been used for farming to improve growth but have not been created in the Kumamoto oyster, Crassostrea sikamea, which is one of the crucial aquaculture species on the southern coast of China. In the present study, triploids were created using cytochalasin B to inhibit polar body II release in C. sikamea, with the untreated oysters as controls. Triploidy rates of 87 and 57.67%, on average, were obtained in larvae and adults, respectively. Larval growth and survival of the triploid were significantly lower than that of the controls (p < 0.05). In contrast, the triploid postlarvae and adults had a significant growth advantage over the controls (p < 0.05) during the period of 180 (December) to 450 days (September of the next year). Moreover, the triploids clearly exhibited significant sterility in the reproductive season. The glycogen and triglyceride contents in the gonad, adductor muscle, mantle, and gill were higher in triploids than in controls from 180 to 450 days. As a result, high physiological energy supply was strongly correlated with superior growth and reduced reproduction in triploid C. sikamea. The triploid C. sikamea is an excellent oyster species and can be used to improve growth for C. sikamea farming.     

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.