Triploid Fish Research Articles

Chromosomal aberrations and early mortality in a non-mammalian vertebrate: example from pressure-induced triploid Atlantic salmon.

In commercial aquaculture, the production of triploid fish is currently the most practical approach to prevent maturation and farm-to-wild introgression following escapes. However, triploids often exhibit poor welfare, and the underlying mechanisms remain unclear. Inheritance issues associated with sub-optimal hydrostatic pressure treatments used to induce triploidy, or the genetic background of parental fish, have been speculated to contribute. We tested this by quantifying the frequency and type of chromosomal aberrations in Atlantic salmon subjected to a gradient of sub-optimal pressure treatments (Experiment 1) and from multiple mothers (Experiment 2). From these experiments, we genotyped a subsample of ~900 eyed eggs and all ~3300 surviving parr across ~20 microsatellites. In contrast to the low frequency of chromosomal aberrations in the diploid (no hydrostatic pressure) and triploid (full 9500 PSI treatment) controls, eyed eggs subjected to sub-optimal pressure treatments (6500-8500 PSI) had a higher incidence of chromosomal aberrations such as aneuploidy and uniparental disomy, corresponding to lower triploidization success and higher egg mortality rates. We also observed maternal effects on triploidization success and incidence of chromosomal aberrations, with certain half-sibling families exhibiting more aberrations than others. Chromosomal aberrations were rare among surviving parr, suggesting a purge of maladapted individuals during early development. This study demonstrates that sub-optimal hydrostatic pressure treatments and maternal effects not only influence the success of triploidization treatments, but may also affect the incidence of chromosomal aberrations and early mortality. The results have important implications for aquaculture breeding programs and their efforts to prevent farm-to-wild introgression.

UBE2J1 suppresses interferon signaling by facilitating the ubiquitination and degradation of IRF7

Ubiquitin-binding enzyme E2J1 (UBE2J1) is a predominant member of the UBE2 family. In mammals, it is involved in antiviral immunity, ER-misfolded protein degradation, and ubiquitination. However, the function of UBE2J1 in teleost has not been elucidated. In the present study, we cloned the homolog of UBE2J1 in triploid hybrid fish and investigated its regulatory role in the antiviral interferon (IFN) signaling. We determined that the full-length coding sequence of the triploid fish UBE2J1 (3nUBE2J1) consists of 939 base pairs and encodes 313 amino acids. Upon SVCV infection, the mRNA levels of 3nUBE2J1 were increased in the spleen, liver, kidney, and the caudal fin cell line of triploid fish. Knockdown of 3nUBE2J1 in host cells and fish enhanced their antiviral response and curtailed SVCV replication. Conversely, overexpression of 3nUBE2J1 inhibited the activation of IFN promoter induced by SVCV, poly (I:C), or RLR pathway factors. Further studies revealed that 3nUBE2J1 interacted with 3nIRF7 and suppressed both the protein level and antiviral activity of 3nIRF7. Finally, we demonstrated that 3nUBE2J1 enhanced the K27- and K29-linked ubiquitination of 3nIRF7, leading to its degradation. Collectively, our findings indicate that triploid fish UBE2J1 serves as a novel negative regulator of IFN signaling by inhibiting 3nIRF7.

Asexuality shapes traits in a hybrid fish

Animal morphology is influenced by several factors, including gonadal development and gametogenesis. Although their effects are well documented in male/female differentiation, much less is known about same-sex effects, such as those caused by their mode of reproduction. Here, using geometric morphometric analyses, we compare two groups of all-female triploid hybrid fish Chrosomus eos × eos-neogaeus, that differ only by their sexual and asexual reproductive strategies. We demonstrate that morphological differences arise from factors inherently associated with their mode of reproduction, with results replicated in two distinct lineages and in natural and common garden environments. Such differences provide additional insight about the costs and benefits of both reproductive strategies, which have mostly been of a demographic, population genetic, or genetic nature. In particular, these findings have important implications for the ecology of asexual organisms and contribute to the study of sex evolution by adding complexity to the paradox of sex theory.

Selective autophagy receptor p62/SQSTM1 inhibits TBK1-IRF7 innate immune pathway in triploid hybrid fish

The production of type I interferon is tightly regulated to prevent excessive immune activation. However, the role of selective autophagy receptor SQSTM1 in this regulation in teleost remains unknown. In this study, we cloned the triploid fish SQSTM1 (3nSQSTM1), which comprises 1371 nucleotides, encoding 457 amino acids. qRT-PCR data revealed that the transcript levels of SQSTM1 in triploid fish were increased both in vivo and in vitro following spring viraemia of carp virus (SVCV) infection. Immunofluorescence analysis confirmed that 3nSQSTM1 was mainly distributed in the cytoplasm. Luciferase reporter assay results showed that 3nSQSTM1 significantly blocked the activation of interferon promoters induced by 3nMDA5, 3nMAVS, 3nTBK1, and 3nIRF7. Co-immunoprecipitation assays further confirmed that 3nSQSTM1 could interact with both 3nTBK1 and 3nIRF7. Moreover, upon co-transfection, 3nSQSTM1 significantly inhibited the antiviral activity mediated by TBK1 and IRF7. Mechanistically, 3nSQSTM1 decreased the TBK1 phosphorylation and its interaction with 3nIRF7, thereby suppressing the subsequent antiviral response. Notably, we discovered that 3nSQSTM1 also interacted with SVCV N and P proteins, and these viral proteins may exploit 3nSQSTM1 to further limit the host's antiviral innate immune responses. In conclusion, our study demonstrates that 3nSQSTM1 plays a pivotal role in negatively regulating the interferon signaling pathway by targeting 3nTBK1 and 3nIRF7.

Diploid and triploid Chinook salmon (Oncorhynchus tshawytscha) exhibit differential immunological responses toacute thermal stress.

Exposure to temperatures outside of a fish's optimal range results in suppression of the immune system, ultimately leaving aquaculture stocks susceptible to disease outbreaks. This effect is exacerbated in triploid fishes, which demonstrate greater susceptibility to stress than their diploid counterparts. This study investigates the impacts of acute heat stress on the abundance of immune transcripts and proteins in diploid and triploid Chinook salmon (Oncorhynchus tshawytscha), an important finfish crop. This study also demonstrates that acute heat stress induces significant increases in the abundance hsp70, hsp90 and il1b transcripts in the head kidneys, gills and heart ventricles of both diploid and triploid Chinook salmon. Widespread dysregulation of antigen-presentation transcripts was also observed in fish of both ploidies. These results suggest that acute heat stress activates acute-phase responses in Chinook salmon and dysregulates antigen presentation, potentially leaving fish more susceptible to infection. At the protein level, IL-1β was differentially expressed in the head kidney and ventricles of diploid and triploid salmon following heat shock. Differential expression of two tapasin-like proteins in diploid and triploid salmon subjected to heat shock was also observed. Altogether, these data indicate that diploid and triploid Chinook salmon respond differently to acute thermal stressors.

Triploid brown trout, Salmo trutta, develop functional gonads with age and are able to interbreed with diploid counterparts.

The study investigated if gonad maturation in triploid brown trout, Salmo trutta, was entirely suppressed or only delayed, and if triploids could interbreed with diploid counterparts. Ten percent of the total number of 3-year-old triploid S. trutta, 15% of 4-year-old fish, and 17% of 5-year-old fish produced semen. Three and 4 years old triploid fish did not produce eggs, but 15% of the 5-year-old fish did so. The quantity and sperm motility of triploid semen did not differ from diploids, but the sperm concentration was significantly lower. When diploid eggs were fertilized with triploid semen (3n × 2n crosses), the percentage of eyed stage embryos, of hatched larvae, and of normal-shaped larvae did not differ from the diploid controls. Circa 90% of 3n × 2n crosses had a ploidy level of 2.4n. In the remaining percentage of 3n × 2n crosses, the ploidy level was ≥2n and <2.4n. In sperm competition experiments where diploid eggs were fertilized with a mixture of diploid and triploid semen, 52% of the originating larvae had a ploidy level of 2n, 43% of 2.4n, and 5% of the fish were not exactly classified. From the start of feeding to an age of 248 days, the mortality rate of 3n × 2n interploid crosses and of 2n × 2n controls was similar. The growth of interploid crosses was significantly higher than that of controls. In triploid mature females, the egg mass per kilogram of body weight was significantly lower than in diploids. The mass of the non-hardened eggs and the percentile weight increase during hardening did not differ from diploid eggs. When triploid eggs were fertilized with diploid semen (2n × 3n crosses), the development rate to normal hatched larvae was less than 10%. All originating larvae had a ploidy level of 3n. From the start of feeding to an age of 248 days, 2n × 3n crosses had a higher mortality rate (15%) than diploid controls (<5%). Growth of this type of interploid crosses was reduced in comparison to controls. Therefore, triploids introduced into natural waters for recreational fisheries or escaping from farms may interbreed with diploid counterparts. This not only alters the genotypes of local populations but also changes the ploidy levels.

Initial investigations into the production of triploid sunshine bass using temperature shock

AbstractObjectiveThe sunshine bass (female White Bass Morone chrysops × male Striped Bass M. saxatilis) is an important food fish raised in U.S. aquaculture. Spawning is performed by manually stripping and fertilizing White Bass eggs with Striped Bass sperm. Embryos hatch in approximately 48 h. Mature sunshine bass females can become fertile and produce viable eggs, which is problematic in commercial food‐fish production; therefore, triploid fish are desired.MethodsThe present nonreplicated study incorporated three warm and three cold temperature shocks, along with three postfertilization initiation times and three exposure times for each temperature, during initial stages of incubation to induce triploidy. After exposure to temperature shocks, fertilized eggs were incubated in experimental hatching systems. The best performing cold and warm temperatures and times (and an ambient diploid control) were then used to incubate fertilized eggs in a commercial‐scale trial using standard industry practices with a single batch of eggs. Larvae were stocked at about 185,000 larvae/ha (75,000 larvae/acre) in newly filled and fertilized ponds. Fry were sampled weekly and harvested at 30 days.ResultThere were 15 survivors from the pond containing fish derived from cold‐shocked fertilized eggs; these individuals were tested via Coulter Counter, and no triploids were detected. Testing of 50 survivors from the pond containing fish derived from warm‐shocked fertilized eggs indicated that 14% were triploid.ConclusionTherefore, initial trials indicate that warm temperature shock is preferable to cold shock for inducing a greater percentage of triploid sunshine bass.

GSTP1 is a negative regulator of MAVS in the antiviral signaling against SVCV infection

Glutathione S-transferase P1 (GSTP1), the most ubiquitous member of the GST superfamily, plays vital roles in the detoxification, antioxidant defense, and modulation of inflammatory responses. However, limited studies have been conducted on the function of GSTP1 in antiviral innate immunity. In this study, we have cloned the homolog of GSTP1 in triploid hybrid crucian carp (3nGSTP1) and investigated its regulatory role in the interferon signaling pathway. The open reading frame of 3nGSTP1 is composed of 627 nucleotides, encoding 209 amino acids. In response to spring viremia of carp virus (SVCV) infection, the mRNA level of 3nGSTP1 was up-regulated in the liver, kidney, and caudal fin cell lines (3 nF C) of triploid fish. The knockdown of 3nGSTP1 in 3 nF C improved host cell's antiviral capacity and attenuated SVCV replication. Additionally, overexpression of 3nGSTP1 inhibited the activation of IFN promoters induced by SVCV infection, poly (I:C) stimulation, or the RLR signaling factors. The co-immunoprecipitation assays further revealed that 3nGSTP1 interacts with 3nMAVS. In addition, 3nGSTP1 dose-dependently inhibited 3nMAVS-mediated antiviral activity and reduced 3nMAVS protein level. Mechanistically, 3nGSTP1 promoted ubiquitin-proteasome degradation of MAVS by promoting its K48-linked polyubiquitination. To conclude, our results indicate that GSTP1 acts as a novel inhibitor of MAVS, which negatively regulates the IFN signaling.

An evaluation of the relative size, body condition, and survival of triploid Walleye in the wild

AbstractObjectiveNatural dispersal or unauthorized introductions of nonnative predatory sport fish can undermine native species conservation and other fisheries management objectives. The use of reproductively sterile triploid fish may provide a lower risk stocking option (i.e., genetic and reproductive containment) for diversifying angling opportunities while possibly reducing angler motivation to illegally transplant fish. Management interest in triploid Walleye Sander vitreus is growing, and these fish are currently stocked by several fisheries agencies. However, little is known about the poststocking growth, body condition, and survival of triploid Walleye—information that is important for calibrating expectations and guiding policy regarding their use.MethodsWe compared the age‐specific size, body condition, and survival of subadult (i.e., ages 0–3) triploid Walleye to those of normal diploid conspecifics in two eastern Colorado reservoirs. Paired stockings of triploid and diploid Walleye occurred over 3 years. Both fry and fingerlings were stocked each year.ResultTriploids were generally smaller in length (6.6% smaller on average) and weight (20.6% smaller) compared to diploids across all age‐classes. Despite being smaller, triploids had similar body condition relative to diploids. On average, the survival of triploids from spring stocking to their first fall was 6.3× lower than that of diploids, but no differences in survival were observed beyond this early life stage. This pattern was consistent across cohorts and reservoirs.ConclusionThe present work provides a foundation for future studies that are designed to clarify patterns of survival from spring to the first fall by accounting for size at stocking and natural‐origin fish and studies that are planned to assess the poststocking performance of triploid Walleye independent of diploids.

Maternal ploidy shapes reproductive pathways in the triploid hybrid Chrosomus eos × eos-neogaeus.

Elements transferred from a mother to her eggs may strongly influence the phenotype of her offspring. Such maternal effects depend on the genotype of the mother, and while multiple ploidy levels occur naturally in some vertebrate species, studies evaluating the impact of maternal ploidy on offspring are scarce. This paper aimed to test whether maternal ploidy is responsible for the two reproductive phenotypes observed in the triploid fish Chrosomus eos × eos-neogaeus. Indeed, these hybrids have two different maternal origins (diploid or triploid) and display two reproductive phenotypes, ameiotic and meiotic hybridogenesis, resulting in diploid and haploid eggs, respectively. To this end, we first conducted a genomic survey to identify epigenetic variations in triploid larvae reared under common garden conditions, concordantly with their maternal origin. The results revealed that the polymorphic epigenetic loci of the larvae clustered into two highly distinct groups consistently with the ploidy of their mother. Diagnostic epigenetic loci were then tested in triploid adult females whose reproductive pathways were already known, to infer their own maternal origin. Altogether, the results suggest that triploid larvae from diploid and triploid mothers will develop the ameiotic and meiotic hybridogenesis pathway, respectively. This confirms that the development of a given reproductive pathway in triploid females results from the ploidy of their mother. Overall, this study supports a strong maternal effect, introducing maternal ploidy and reproductive pathways as additional cause and effect of maternal effects, respectively.

Production of sterile mono-sex triploid yellow drum (Nibea albiflora): genotypic females and sex-reversed phenotypic males with emphasis on utilization as surrogate broodstock.

Production of sterile mono-sex fish is of great significance for sustainable aquaculture as well as germ cell transplantation. In this study, we aimed to produce mono-sex triploid yellow drum, including genotypic females (XXX female) and sex-reversed phenotypic males (XXX male). Firstly, the mono-female triploids were produced through cold-shock treatment on eggs fertilized with sperm from neo-males. Then, the mono-male triploids were produced by the sex reversal of mono-female triploids with oral administration of letrozole (LZ). We comparatively investigated the growth and gonadal development in the mono-sex triploids. The results showed that the triploids displayed similar growth performance to their diploids throughout their first year, but had impaired gonadosomatic index and gametogenesis. No mature gametes were produced in the triploids during their first spawning season. Meanwhile, we analyzed the process of gametogenesis in the both sex of triploids. Ultrastructure of gametogenesis showed that the germ cells arrested at abnormal metaphase 1 in females, while males had irregular meiotic divisions, variable-sized spermatid and degenerated cells. The expression levels of meiosis-related genes (i.e., sycp3 and rec8) confirmed the abnormal meiosis in the triploids. Furthermore, the gonadal development was also determined by the expression patterns of vasa, dmrt1 and cyp19a1a. Abnormal expression of vasa mRNA and protein were detected in triploids. High cyp19a1a expression levels suggested the sex steroid hormones production might be at least partially functional in triploid females. In addition, high dmrt1 expression levels confirmed the masculinization and testicular development of sex-reversed triploid males by LZ. Our findings provide an efficient protocol to produce sterile mono-sex triploid yellow drum and provide new insights into the mechanism of gonadal sterility of triploid fish.

Diploid and triploid Chinook salmon (Oncorhynchus tshawytscha) have altered microRNA responses in immune tissues after infection with Vibrio anguillarum

Production of sterile fishes through artificial retention of a third set of chromosomes (triploidy) is a sustainable alternative for aquaculture since it reduces escapee pressure on wild populations. However, these fishes have reduced survival in stressful conditions and in response to infection. In this study, the impact of Vibrio anguillarum infection on diploid and triploid Chinook salmon (Oncorhynchus tshawytscha) was investigated to identify if there was any significant immune regulation by microRNAs (miRNA). Small RNAs from hindgut, head kidney, and spleen were sequenced to determine if miRNA transcript abundance was altered due to ploidy and infection in nine-month old full-sibling diploids and triploids. All three tissues had differentially expressed miRNA prior to infection, indicating subtle changes in epigenetic regulation due to increased ploidy. Additionally, miRNA were altered by infection, but there was only a difference in spleen miRNA expression between diploids and triploids at three days of infection. Furthermore, one miRNA (ssa-miR-2188-3p) was confirmed as having an altered response to infection in triploids compared to diploids, implicating potential immune dysregulation due to increased ploidy. The miRNAs identified in this study are predicted to target immune pathways, providing evidence for their importance in regulating responses to pathogens. This study is the first to investigate how increased ploidy alters miRNA expression in response to infection. Additionally, it provides evidence for epigenetic dysregulation in triploid fishes, which may contribute to their poor performance in response to stress.

HnRNPM suppressed IRF7-mediated IFN signaling in the antiviral innate immunity in triploid hybrid fish

Mammalian heterogeneous nuclear ribonucleoproteins M (hnRNPM) is a critical splicing regulatory protein that has been reported to negatively regulate the RLR signaling pathway by impairing the binding of RIG-I and MDA5 to viral RNA. To explore the role of hnRNPM in the antiviral innate immune response in teleost fish, the hnRNPM homologue of triploid fish (3nhnRNPM) has been cloned and identified in this paper. The CDS of 3nhnRNPM gene is composed of 2016 nucleotides and encodes 671 amino acids. 3nhnRNPM migrated around 71 kDa in immunoblotting assay and was mainly detected in the nucleus in nucleo-cytoplasmic separation assay and immunofluorescent staining test. When 3nhnRNPM and 3nIRF7 were co-expressed in EPC cells, 3nhnRNPM significantly reduced the 3nIRF7-induced interferon (IFN) promoter transcription. Correspondingly, the mRNA levels of the SVCV-M, -N, -P, and -G genes were noteworthily enhanced, but the transcription levels of epcIFNφ1, epcMx1, epcPKR, and epcISG15 were dramatically decreased. Additionally, the knockdown of 3nhnRNPM resulted in restricted SVCV replication and enhanced host cell antiviral activity. Furthermore, the association between 3nhnRNPM and 3nIRF7 has been identified by the co-immunoprecipitation assay. In addition, we found that 3nIRF7 was detained in the nucleus when co-expressed with 3nhnRNPM. To sum up, our data supported the conclusion that 3nhnRNPM suppressed 3nIRF7-mediated IFN signaling in the antiviral innate immunity.

Telomerase Activity in Somatic Tissues and Ovaries of Diploid and Triploid Rainbow Trout (Oncorhynchus mykiss) Females.

Telomerase activity has been found in the somatic tissues of rainbow trout. The enzyme is essential for maintaining telomere length but also assures homeostasis of the fish organs, playing an important role during tissue regeneration. The unique morphological and physiological characteristics of triploid rainbow trout, when compared to diploid specimens, make them a promising model for studies concerning telomerase activity. Thus, in this study, we examined the expression of the Tert gene in various organs of subadult and adult diploid and triploid rainbow trout females. Upregulated Tert mRNA transcription was observed in all the examined somatic tissues sampled from the triploid fish when compared to diploid individuals. Contrastingly, Tert expression in the ovaries was significantly decreased in the triploid specimens. Within the diploids, the highest expression of Tert was observed in the liver and in the ovaries of the subadult individuals. In the triploids, Tert expression was increased in the somatic tissues, while the ovaries exhibited lower activity of telomerase compared to other organs and decreased compared to the ovaries in the diploids. The ovaries of triploid individuals were underdeveloped, consisting of only a few oocytes. The lack of germ cells, which are usually characterized by high Tert expression, might be responsible for the decrease in telomerase activity in the triploid ovaries. The increase in Tert expression in triploid somatic tissues suggests that they require higher telomerase activity to cope with environmental stress and maintain internal homeostasis.

Gene expression of Lysozyme C and Lysozyme G in diploid and triploid Astyanax lacustris experimentally infected with Aeromonas jandaei

Triploid fish are becoming increasingly popular in commercial aquaculture due to their sterility, which results in improved meat quality and better growth rates as a result of the lack of sexual maturation. However, triploid fish are generally perceived as being more susceptible to diseases and less able to cope with suboptimal environmental conditions. The present study aimed to evaluate the effects of triploidy on the expression of C-type and G-type lysozymes (LysC and LysG) in lambaris (Astyanax lacustris) that were challenged with a pathogenic strain of Aeromonas jandaei. The animals were induced to triploidy as described in the literature. We infected 10 diploid and 10 triploid fish with 104 CFU of A. jandaei in 0.1 mL of PBS by intraperitoneal injection, a dose capable of triggering an immune response in the animals. Fish from the control groups were inoculated with 0.1 mL of sterile PBS. Molecular cloning of Lysozyme C and Lysozyme G was performed to characterize these genes, and the analysis of gene expression for LysC and LysG of A. lacustris diploids and triploids was conducted using qPCR. The collected data revealed that there were no significant differences in basal expression between diploid and triploid uninfected fish. However, the challenged fish of both ploidies exhibited an increase in expression compared to the control group. The results of study indicate that infected diploid fish exhibit significantly higher lysozyme expression in the spleen compared to infected triploid fish. While basal lysozyme expression levels between diploid and triploid fish remain unaffected, our findings suggest that triploidy may lower lysozyme expression in response to infection, with diploid fish exhibiting higher levels of expression.

Endurance Swimming Is Related to Summer Lake Survival of Rainbow Trout in a Warm Lake with Avian Piscivores

Fitness of fish is assumed to be influenced by locomotion performance, but empirical evidence linking swimming capacity to survival in nature remains sparse. Poor triploid (3N) fish aerobic swimming performance in conjunction with production of sibling diploid (2N) and 3N populations of genetically identical origin to minimize variability among compared populations make 3N trout an informative system to test hypotheses about fitness consequences of fish locomotion. Here, we ask if reduced survival of 3N relative to 2N trout in natural ecosystems during periods of high temperature relate to a lower aerobic swimming capacity and aerobic scope of 3N compared with 2N conspecifics. Three-yearly cohorts of conspecific 2N and 3N hatchery-reared, yearling rainbow trout were ranked for swimming endurance, externally marked for their endurance quantile, and then stocked into two lakes as yearlings to quantify their survival in the wild over summer as a function of ploidy, temperature and endurance; all while tracking temperature and depth habitat utilization via telemetry. As expected, 3N swimming endurance was lower than that of 2N, but with considerable individual overlap. Aerobic swimming endurance, especially for 3N, was predictive of summer survival in a warm lake where piscivorous birds potentially exerted high predation pressure, resulting in low fish survival. This empirical evidence of a connection between swimming endurance and fitness provides support for long held assumptions of this relationship and could inform future sport fishing stocking industry practices to match fish strains to ideal habitats.

Comparative study on the fillet nutritional quality of diploid and triploid rainbow trout (Oncorhynchus mykiss)

This study was conducted to investigate the fillet nutritional quality difference of diploid and triploid rainbow trout with the same genetic background, farming condition and body weight. The results showed that, 1) diploid rainbow trout had significantly higher content of protein, valine, cysteine, histidine and proline, and higher essential amino acid index, nutrition index and biological value than triploid rainbow trout (P < 0.05). However, essential amino acids and total amino acids contents as well as amino acid score and chemistry score were similar between the two fish (P > 0.05), which indicated that both diploid and triploid rainbow trout fillets were balanced and suitable for human consumption. 2) Triploid rainbow trout had significantly higher content of lipid, C18:1n-9, C18:2n-6, C20:5n-3, C22:6n-3, unsaturated fatty acids (UFA), polyunsaturated fatty acids (PUFA), long chain polyunsaturated fatty acids, n-3 series fatty acids, n-6 series fatty acids and total fatty acids (TFA) than diploid rainbow trout (P < 0.05). However, no differences were found in saturated fatty acids (SFA), C18:3n-3 and C20:4n-6 contents between the two fish (P > 0.05). Based on PUFA/SFA, UFA/SFA, PUFA/TFA, atherogenic index, and thrombogenic index, both diploid and triploid rainbow trout consumption were beneficial for human health, triploid fish had an even greater advantage. 3) Triploid rainbow trout had higher content of sodium, calcium, iron, manganese and zinc but lower content of magnesium, copper and selenium than diploid rainbow trout (P < 0.05). Multivariate statistical analysis of 79 quality indicators also showed that the fillet nutritional composition of triploid rainbow trout was different with its diploid counterparts. The reason may be related to the difference of lipid metabolism in rainbow trout with different ploidy.

Comparative study on the gonadal development in the diploid and artificially induced triploid olive flounder Paralichthys olivaceus

Artificial induction and breeding of triploids has been widely performed in fish due to their sterility or low fecundity. However, detailed studies on the gonadal development status, gamete formation, fertilization ability, and molecular mechanism of the induced triploids in marine fish are limited. Olive flounder Paralichthys olivaceus is an important mariculture fish in China, Japan, and Korea. In this study, by observing the gonadal development of the artificially induced triploid and control diploid flounder, the histological structure, sex hormone level, and transcriptome profiles were compared. The fecundity of the male triploid fish was also evaluated. The results showed that the development of the gonads of the triploids was severely inhibited. Apoptosis occurred in both male and female triploid gonads, which may lead to inhibition of germ cell development. At 24 months post hatch (mph), the contents of testosterone (T) and estradiol (E2) in the gonads of the female triploid flounder were lower than those of the diploid ones (P < 0.05), and the contents of E2 in the testes of the male triploid flounder were higher than those of the diploid ones (P < 0.05). Although the triploid flounder could produce sperm, the sperm presented different sizes of the head, deletion of the mitochondrion, and short tail under the scanning electron microscope. Only a few spermatozoa were slightly active. The ploidy of the spermatozoa included haploid, haploid - diploid, triploid, 4-5n aneuploidy, and even higher ploidy. By using sperm from the triploids with the control eggs, very few abnormal embryos and larvae were obtained. Transcriptome data at 18 and 24 mph showed that most of the significantly changed differentially expressed genes between the diploids and triploids were related to cell division and proliferation, energy and metabolism, fertility, and apoptosis. In the triploid testis, many genes related to spermatogenesis, such as mns1, dnah1, and dnah5, were down-regulated; while in the triploid ovary, genes related to testicular development, such as amh, amhr2, and wnt4, were up-regulated, which also provided molecular evidence for the testicular-like gonads observed in the histology of the triploid ovary. These results may help explain the developmental retardation and low fecundity in the gonads of triploid fish that are important for the future application of culture.

Induction of high triploid rate blunt snout bream (Megalobrama amblycephala) by hydrostatic pressure and analysis of gonadal sterility mechanism

At present, only cold shock method was used to induce the triploid of M. amblycephala, and because the triploid rate is not high enough, the triploid fish has not been applied in aquaculture on a large scale. This study aimed to obtain the best induction conditions of triploid by hydrostatic pressure, reveal the reproductive characteristics and mechanism of gonadal sterility of triploid M. amblycephala, provide an insights into the meiotic patterns of triploid fish. We successfully induced triploid in M. amblycephala using the hydrostatic pressure method. The highest triploid rate reached 85.92% at 55 MPa for 3 min after 3 min of fertilization. The relative DNA content of the triploid fish was 1.5 times of that of the diploid fish, and their chromosome karyotype was 3n = 72 = 27 m + 39sm + 6st. Both female and male triploids had dysplastic gonads, the testis of triploid males was able to produce 1.5n sperm without fertility because all the hybrid embryos of 2n♀ × 3n♂ died; however, the ovaries of triploid females belonged to complete sterility. The verification of gene expression patterns and interaction suggested that the hypothalamus-pituitary-gonad (HPG) axis process was disturbed and unsynapsed chromosome (UC) axis was blocked may jointly caused the complete sterility without egg production of female triploids. However, triploid males belonged to uncompletely sterility, which can produce a small number of sperm and complete fertilization, but the offspring cannot survive, may caused by the not completely blocked UC axis with low expression of hormad1 and iho1.

Origin of scarlet gynogenetic triploid Carassius fish: Implications for conservation of the sexual-gynogenetic complex.

Conservation of sperm-dependent asexual (gynogenetic) species is challenging due to their complicated ecological dynamics, which requires the stable coexistence with their sperm-providing sexual relatives, who often share similar niches. A symbolic but vulnerable gynogenetic animal is the scarlet Carassius fish, or Hibuna, which is mainly found in Lake Harutori on Hokkaido, Japan. Although Hibuna in Lake Harutori has been protected as a symbol of the Natural Monument of Japan, it has recently suffered population decline. To establish an effective conservation strategy for Hibuna, we investigated its origin, reproductive mode, and genetic diversity, with reference to the surrounding wild populations, using nuclear microsatellites and mitochondrial gene sequences. Our genetic analyses revealed that the main ploidy of Hibuna was triploid or tetraploid, and it reproduces gynogenetically. However, no co-existing sexual diploid Carassius was detected among our samples, suggesting that the sexual diploids and the gynogenetic population including Hibuna would be at risk of co-extirpation. In addition, Hibuna showed high genetic/clonal diversity and most Hibuna had nonindigenous mitochondrial haplotypes that are mostly identical to those reported from goldfish. These results indicate that Hibuna most probably originated from hybridization between indigenous gynogenetic triploids and goldfish introduced about 100 years ago, involving rare sexual reproduction. This spontaneous long-term field experiment exemplifies the recently documented diversification process of gynogenetic Carassius via complex interploidy gene flow. Although the priority to be placed on the conservation of Hibuna is controversial, the maintenance of gynogenetic Carassius, including Hibuna, requires strategic conservation of sexual populations.