All-female Fish Research Articles

Study on hormone induction of the male parent and embryonic development, gonad differentiation, and growth of “All-female No.1” Culteralburnus

Female Culter alburnus was faster in growth rate than males. In this study, the gynogenetic G2 and the pseudo-male G2' were used as the female and male parents, respectively, to breed a new national variety “All-female No.1” C. alburnus (AFC). Hormone induction, embryonic development, gonadal differentiation, and growth of AFC were studied. The results showed induction with low concentrations of 17α-methyltestosterone in a indoor-net cage culture was not effective. Under the stimulation of 17α-methyltestosterone, some gonads had a tendency to transform into testis, but not completely. There were three types of gonads in 5-month-old and four types of gonads in 12-month-old fishes, however, they all differentiated into ovaries in 15-month-old fishes. Testosterone propionate and high concentrations of 17α-methyltestosterone in pond culture induction had a good effect resulting in ①a functional pseudo-male with normal testis development that could successfully extrude semen during the breeding period, ②a pseudo-male with normal testis development, but could not extrude semen, and ③the appearance of intersexual glands. The second experiment revealed that with common fish, all-female fish embryo had normal embryonic development. The development time and morphological characteristics of each stage were similar, but the development of the all-female embryo was slightly slower than the common embryos. The gonad differentiation of the all-female embryo were normal and none differentiated into testis, which indicated that all-female could ensure the female sex without affecting the normal gonad differentiation. The correlation between body weight, length, and month-age of all-female and common fish was strong. The all-female had faster growth rate and more uniform growth specification than the common fish. Therefore, the use of testosterone propionate and high concentrations of 17α-methyltestosterone in pond culture induction could avoid complete degeneration of gonads into ovaries. The all-female embryo had the advantages of normal embryonic development and gonadal differentiation, faster growth, and uniform growth specification.

Molecular and Physiological Effects of 17α-methyltestosterone on Sex Differentiation of Black Rockfish, Sebastes schlegelii.

It is widely known that all-female fish production holds economic value for aquaculture. Sebastes schlegelii, a preeminent economic species, exhibits a sex dimorphism, with females surpassing males in growth. In this regard, achieving all-female black rockfish production could significantly enhance breeding profitability. In this study, we utilized the widely used male sex-regulating hormone, 17α-methyltestosterone (MT) at three different concentrations (20, 40, and 60 ppm), to produce pseudomales of S. schlegelii for subsequent all-female offspring breeding. Long-term MT administration severely inhibits the growth of S. schlegelii, while short term had no significant impact. Histological analysis confirmed sex reversal at all MT concentrations; however, both medium and higher MT concentrations impaired testis development. MT also influenced sex steroid hormone levels in pseudomales, suppressing E2 while increasing T and 11-KT levels. In addition, a transcriptome analysis revealed that MT down-regulated ovarian-related genes (cyp19a1a and foxl2) while up-regulating male-related genes (amh) in pseudomales. Furthermore, MT modulated the TGF-β signaling and steroid hormone biosynthesis pathways, indicating its crucial role in S. schlegelii sex differentiation. Therefore, the current study provides a method for achieving sexual reversal using MT in S. schlegelii and offers an initial insight into the underlying mechanism of sexual reversal in this species.

Reproductive Cycle, Size at Maturity and Fecundity of Giuris margaritaceus in Limboto Lake

Graphical Abstract Highlight Research The size of the first mature gonads in hulu'u fish can be analyzed. Differences in sperm and egg size characterize sexual maturity. Sexual maturity is not influenced by season. Hulu'u fish have an asynchronous spawning type. Abstrack Understanding fish reproduction biology, particularly sexual maturity, the reproductive cycle, and fecundity, is useful for managing fisheries. The reproduction of Giuris margaritaceus in Limboto Lake is regrettably unknown. Over a year, fish specimens (N = 662) were gathered. They ranged in length from 6.5 to 20.9 cm (mean SD 10.9±2.5). Fecundity varied from 30,057 to 61,920. The first sexual maturity of male and female fish occurred at different sizes, 11.2 cm and 10.3 cm, respectively. This information is needed to determine this fish's lowest acceptable harvest length under the management aim to enter all-female fish for reproduction at least once. We estimated the reproductive of G. margaritaceus cycle using the size of the oocytes, histological analysis of the ovaries, and monthly mean GSI. These techniques produced the same reliable conclusion: G. margaritaceus can spawn yearly, with maximal between September and October. Based on this finding, spawners of Giuris margaritaceus can be kept from entering the reproductive stock between September and October. Future efforts to fine-tune fishing efforts for the effective management of G. margaritaceus may be aided by the study's findings.

Effective “off-on” switch for fertility control in female zebrafish

The implementation of a controllable sterility strategy is crucial for the commercialization of precise trait improvements in farmed fish using genome editing and sustainable development of fisheries. Our previous research has demonstrated that females deficient in pituitary gonadotropin luteinizing hormone β-subunit (lhβ) or gonadal steroidogenesis gene steroidogenic acute regulatory protein (star) exhibit sterility due to impaired oocyte maturation and ovulation. Nevertheless, the effective restoration of fertility in lhβ- or star-deficient females remains unsolved. This study has discovered that the administration of exogenous 17α,20β-dihydroxy-4-pregnen-3-one (DHP) at 100 and 300 μg/L for 6 h (from 02:00 to 08:00 a.m.) effectively restores the fertility of lhβ- or star-deficient females. Fertilized eggs from these mutant females can be raised without noticeable developmental defects for up to 3 weeks post-fertilization (wpf) compared to the wild-type (WT) control zebrafish. The increased expression levels of adamts9 and adam8b in lhβ- or star-deficient zebrafish females treated with DHP demonstrate a positive correlation with oocyte maturation and ovulation restoration. In contrast, exogenous DHP administration did not rescue the sterility phenotype observed in progesterone receptor (pgr)-deficient females. Building on our recent success in generating an all-female carp population through cytochrome P450, family 17, subfamily A, polypeptide 1 (cyp17a1)-depletion, our research presents a promising and effective strategy for an “off-on” switch for managing fertility in genome-edited cyprinids. The strategy would offer practical guidance and theoretical justification for developing “controllable fertility” in all-female fish, which would support the sustainable development of fisheries by promoting the use of novel biotechnologies in aquaculture in an eco-friendly manner.

Preliminary Analysis of Quality and Cryopreservation of Sex-Reversed Female Culter alburnus Sperm

Sex-reversed females are the fathers of all-female fish and their sperm quality directly affects the quality of all-female fish. In this study, fecundity, physiological characteristics, biochemical characteristics, ultrastructure, and viability of sex-reversed female sperm after cryopreservation were investigated to understand sperm quality. (1) The sex-reversed female sperm had a lower fertilization rate, hatching rate, and larval survival rate of 72 hr and higher larval deformation rate compared with normal male sperm. (2) The changes in activation rate between normal male and sex-reversed female sperm under different salinity and pH conditions were the same; the optimum salinity was between 0 and 4%, and the optimum pH was 5.5. (3) The preservation time of sex-reversed female sperm is significantly shorter than that of normal male sperm at room temperature. (4) Sex-reversed female sperm had lower ATPase content and c/p value than normal male sperm. (5) Ultrastructural injury including damaged membranes of the head and mitochondria, broken flagella, greater mitochondrial damage, and mitochondrial membrane expansion were found in the sex-reversed female sperm. (6) The most suitable antifreeze for cryopreservation of sex-reversed female and normal male sperm was 10% ethylene glycol and dilution was D-15, but the activation rate, movement, and life span of sex-reversed female sperm were reduced after cryopreservation. It was speculated that the sperm mitochondria of the sex-reversed female were more damaged than those of the normal male, which led to differences in fertility and cryopreservation.

Exploring Sexual Dimorphism in the Intestinal Microbiota of the Yellow Drum (Nibea albiflora, Sciaenidae).

Most of fish species exhibit striking sexual dimorphism, particularly during growth. There are also sexual dimorphisms of internal organs and biological functions, including those of intestinal microbiota, which likely plays a key role in growth. In this study, the growth and intestinal microbiota of the female, male, and all-female Nibea albiflora (yellow drums) were comprehensively analyzed. The caged culture female and all-female yellow drums showed higher growth rates than males. A further analysis of the intestinal microbiota showed a significant difference in diversity between females and males in the summer, whereas there were no significant differences in the diversity and richness between females and males in the winter. In contrast, a significant difference in richness was observed between all-female and male fish, regardless of the season. Although the main composition of the intestinal microbiota showed no significant sex differences, the community structure of the intestinal microbiota of yellow drums did. Furthermore, the correlations between intestinal microbial communities are likely to be influenced by sex. The ecological processes of the intestinal microbial communities of the yellow drums showed clear sexual dimorphism. Further network analysis revealed that, although the main components of the network in the intestinal microbiota of female, male, and all-female fish were similar, the network structures showed significant sex differences. The negative interactions among microbial species were the dominant relationships in the intestinal ecosystem, and Bacteroidetes, Firmicutes, and Proteobacteria were identified as the functional keystone microbes. In addition, the functional pathways in the intestinal microbiota of yellow drums showed no significant sexual or seasonal differences. Based on the findings of this study, we gain a comprehensive understanding of the interactions between sex, growth, and intestinal microbiota in yellow drums.

Characteristics and Cryopreservation of Semen of Sex-Reversed Females of Salmonid Fish.

Sex reversal has been used as a breeding strategy by salmonid fish to produce genetically and phenotypically single sex populations. Production of all-female fish has great importance for the creation of monosex female triploids of salmonid fish, which are valued for their sterility, lack of female maturation, and larger commercial size. Among salmonids, the majority of rainbow trout (Oncorhynchus mykiss) production is based on all-female production with a high proportion of all-female triploid production in Europe. The main aim of this review is to present the recent knowledge regarding sex-reversed females (SRFs) of salmonid fish. We discuss the methods of sex reversal as well as their effects on the morphology and histology of the reproductive tract. We focus on the characteristics of SRF semen as well as the factors determining semen quality. The lower quality of SRF sperm compared to that of normal males has resulted in the need for the artificial maturation of semen. Most importantly, methods of semen storage—both short-term and long-term (cryopreservation)—that can improve hatchery operations are presented with the special emphasis on recent progress in development of efficient cryopreservation procedures and use of cryopreserved semen in hatchery practice. Moreover, we also address the emerging knowledge concerning the proteomic investigations of salmonid sperm, focusing primarily on the proteomic comparison of normal male and SRF testicular semen and presenting changes in SRF rainbow trout sperm proteome after in vitro incubation in artificial seminal plasma.

An Investigation of ZZ/ZW and XX/XY Sex Determination Systems in North African Catfish (Clarias gariepinus, ).

An investigation of sex-specific loci may provide important insights into fish sex determination strategies. This may be useful for biotechnological purposes, for example, to produce all-male or all-female fish for commercial breeding. The North African catfish species, Clarias gariepinus, has been widely adopted for aquaculture because its superior growth and disease resistance render the species suitable for hybridization with other catfish to improve the productivity and quality of fish meat. This species has either a ZZ/ZW or XX/XY sex determination system. Here, we investigate and characterize these systems using high-throughput genome complexity reduction sequencing as Diversity Arrays Technology. This approach was effective in identifying moderately sex-linked loci with both single-nucleotide polymorphisms (SNPs) and restriction fragment presence/absence (PA) markers in 30 perfectly sexed individuals of C. gariepinus. However, SNPs based markers were not found in this study. In total, 41 loci met the criteria for being moderately male-linked (with male vs. female ratios 80:20 and 70:30), while 25 loci were found to be moderately linked to female sex. No strictly male- or female-linked loci were detected. Seven moderately male-linked loci were partially homologous to some classes of transposable elements and three moderately male-linked loci were partially homologous to functional genes. Our data showed that the male heterogametic XX/XY sex determination system should co-exist with the ZZ/ZW system in C. gariepinus. Our finding of the co-existence of XX/XY and ZZ/ZW systems can be applied to benefit commercial breeding of this species in Thailand. This approach using moderately sex-linked loci provides a solid baseline for revealing sex determination mechanisms and identify potential sex determination regions in catfish, allowing further investigation of genetic improvements in breeding programs.

Production of market-size European strain Atlantic salmon (Salmo salar) in land-based freshwater closed containment aquaculture systems

Interest in land-based farms using recirculating aquaculture systems (RAS) for market-size Atlantic salmon (Salmo salar) continues to grow, and several commercial facilities are already rearing fish. Performance data for commercially available mixed-sex, all-female, and triploid all-female Atlantic salmon reared to market-size in freshwater land-based facilities, however, are limited, particularly for European strain fish. Accordingly, eight groups of European-sourced Atlantic salmon (five groups of diploid mixed-sex, two groups of diploid all-female, and one group of triploid all-female fish) were reared from eyed egg to market-size in a semi-commercial scale land-based aquaculture systems over five separate production cycles to quantify performance metrics. Fish reached market-size (4−5 kg) in 24.7–26.3 months post-hatch. Fish were reared at a mean water temperature of 12.3–13.7 °C from first feeding to a mean size of 466–1265 g, then 13.3–15.1 °C during growout. On average, all-female groups grew faster than mixed-sex groups; however, environmental conditions and performance of individual cohorts varied. In a comingled production cycle, diploid all-female salmon grew faster than triploid counterparts. Early maturation rates ranged from 0 % to 67 %, with a mean maturation rate of 34 % for diploid mixed-sex fish and 67 % and 13 % for two diploid all-female groups, respectively. Triploid all-female Atlantic salmon did not mature. This research confirms biological and technological feasibility of growing Atlantic salmon to market-size in land-based systems but controlling early maturation of diploid salmon remained a challenge under the conditions utilized in these trials. This research provides important data inputs to optimize operational and financial projections for existing and potential land-based Atlantic salmon farms.

Smaller rival males do not affect male mate choice or cortisol but do affect 11-ketotestosterone in a unisexual-bisexual mating complex of fish

Male mate discrimination may be affected by the social environment (presence or absence of rival males or mates), which can also affect stress and sex hormones (e.g., cortisol and 11-ketotestosterone (11-KT)). The Amazon molly, Poecilia formosa, is an all-female fish species dependent on sperm from mating with male P. latipinna. We investigated male mate choice in P. latipinna between conspecific females and P. formosa with a rival male present and no rival male present. We measured cortisol and 11-KT release rates from all fish. The presence of a rival male had no effect on male mate choice for conspecific females nor overall mating effort. Male 11-KT decreased on the second day after exposure to a rival male on the first day. Focal male 11-KT is positively correlated with the size of the rival male. Both conspecific and heterospecific females released more 11-KT when in the rival male treatment than when not. Neither male nor female cortisol was affected by the presence or absence of the rival male. We did not find an effect of rival males on male mate choice in contrast to our prediction. Instead, our findings may indicate a hormonal response to social competition.

Identification of sex‐specific markers and heterogametic XX/XY sex determination system by 2b‐RAD sequencing in redtail catfish (Mystus wyckioides)

Sex-specific markers provide significant molecular basis for sex control breeding biotechnology to produce all-male or all-female fish in commercial breeding. Redtail catfish (Mystus wyckioides), one of the commercial bagrid catfishes distributed in Southeast Asian, which have a long sexual maturation period that can last 3–5 years and males have apparent growth advantage over females, but its sex determination system remains unknown. In this study, we first applied 2b-RAD-seq approach to identify three male-specific 2b-RAD-tags and one male heterogametic SNP locus and validated by blast to the genome survey sequences and PCR amplification in both wild and breeding populations. To get longer sex-specific region, we performed genome walking and obtained a 4,630 bp of Y-specific sequence and 4,581 bp of X-specific sequence from the 2b-RAD-tag ref189950 with 92.19% nucleotide identity between them. And 9,923 bp/3,935 bp of Y-specific sequences and 8,491 bp/5,172 bp of X-specific sequences were also identified with 77.49% and 57.07% nucleotide identity in ref208528 and ref210837, respectively. Subsequently, three different kinds of sex-specific primers with different length products were designed based on the detected highly sex differentiated regions and could be used to distinguish males and females both in wild and artificially bred populations. What is more, the X-specific fragment was discovered to produce the dosage effect association in females and in males. The data suggest that male heterogametic XX/XY sex determination system should exist in the redtail catfish. More significantly, the sex-specific markers are of great value to protect wild resources and improve the efficiency of all-male breeding practices for aquaculture in the redtail catfish.

Gender-specific differences in gene expression profiles in gynogenetic Pengze crucian carp

Gynogenesis is a form of asexual reproduction that is used to obtain all-female fish stocks. In this study, we were interested in studying gender-specific differences in gene expression profiles in gynogenetic teleosts, using a carp species. The four-month old gynogenetic Pengze crucian carp F1 (Carassius auratus var. pengzensis, Pcc) showed a high ratio of males under laboratory culture condition. The present study aimed to investigate the differences between males and females. The gonadosomatic index of the females was significantly higher than that of the males. Moreover, the hepatosomatic index of the females was significantly lower than that of the males. Vitellogenin B mRNA was abnormally highly expressed in male hepatopancreas and testes compared to females. Similarly, zona pellucida 2 expressed at a significantly high level in the testes. For the sex related genes, dosage-sensitive sex reversal, adrenal hypoplasia congenital critical region on the X-chromosome gene 1, doublesex and mab-3 related transcription factor 1a, nuclear receptor subfamily 5, group A, member 1b and SRY-box containing gene 9a had significantly higher expression levels in the males than in the females, whereas there was no difference in expression of anti-Müllerian hormone, cytochrome P450 family 19 subfamily A member 1A and forkhead box L2 transcripts between the two genders. The females showed higher levels of estrogen but no significant difference in testosterone compared to the males. The data suggest remarkable differences between the two genders of the Pengze crucian carp.

Complete depletion of primordial germ cells in an All-female fish leads to Sex-biased gene expression alteration and sterile All-male occurrence

BackgroundGynogenesis is one of unisexual reproduction modes in vertebrates, and produces all-female individuals with identical genetic background. In sexual reproduction vertebrates, the roles of primordial germ cells on sexual dimorphism and gonadal differentiation have been largely studied, and two distinct functional models have been proposed. However, the role of primordial germ cells remains unknown in unisexual animals, and it is also unclear whether the functional models in sexual reproduction animals are common in unisexual animals.ResultsTo solve these puzzles, we attempt to utilize the gynogenetic superiority of polyploid Carassius gibelio to create a complete germ cell-depleted gonad model by a similar morpholino-mediated knockdown approach used in other examined sexual reproduction fishes. Through the germ cell-depleted gonad model, we have performed comprehensive and comparative transcriptome analysis, and revealed a complete alteration of sex-biased gene expression. Moreover, the expression alteration leads to up-regulation of testis-biased genes and down-regulation of ovary-biased genes, and results in the occurrence of sterile all-males with testis-like gonads and secondary sex characteristics in the germ cell-depleted gynogenetic Carassius gibelio.ConclusionsOur current results have demonstrated that unisexual gynogenetic embryos remain keeping male sex determination information in the genome, and the complete depletion of primordial germ cells in the all-female fish leads to sex-biased gene expression alteration and sterile all-male occurrence.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-2130-z) contains supplementary material, which is available to authorized users.

Isolation of a Cancer-Associated Microchromosome in the Sperm-Dependent Parthenogen Poecilia formosa

In the asexual all-female fish species Poecilia formosa, the Amazon molly, supernumerary chromosomes have frequently been found in both laboratory-reared and wild-caught individuals. While wild-caught individuals with B chromosomes are phenotypically indifferent from conspecifics, individuals carrying B chromosomes from recent introgression events in the laboratory show phenotypic changes. Former analyses showed that the expression of a pigment cell locus is associated with the presence of these B chromosomes. In addition, they contain a so far unidentified locus that confers a higher susceptibility to tumor formation in the presence of pigmentation pattern. Isolation by microdissection and hybridization to metaphase chromosomes revealed that they contain one or several sequences with similarity to a highly repetitive pericentromeric and subtelomeric sequence in A chromosomes. Isolation of one particular sequence by AFLP showed that the B chromosomes contain at least 1 copy of an A-chromosomal region which is highly conserved in the whole genus Poecilia, i.e. more than 5 million years old. We propose it to be a single copy sequence.

Prudent males, group adaptation, and the tragedy of the commons

For almost five decades three threads have coexisted in the evolutionary and ecological literature, with their links only recently becoming visible and some of them still not properly addressed. These are the levels of selection debate, the metaphor of the tragedy of the commons, and the evolutionary study of sexual conflict. We analyze the eco-evolutionary dynamics of a curious system where an asexual all-female fish species (the Amazon molly Poecilia formosa) requires sperm from other species as a developmental trigger, without utilizing the genes from sperm. The dynamics of such a system bear strong resemblance to host–parasite dynamics, and populations of the sexual ‘host’ species persist much better if males avoid mating with Amazons. However, such avoidance may compromise their current mating success, and if this is the case, prudent mating becomes an altruistic trait that helps to keep an accumulating problem of a competing species at bay, and Amazon-free space can be seen to form a common good that a population should maintain for future generations. A model shows that the evolution of altruistic mating restraint is possible but selection for short-term gains means that it will remain less than perfect. This helps to explain why the anomalous gynogenetic system can persist, but it also raises questions about what kinds of traits can be classified as adaptations when optimization is not perfect and traits evolve to achieve short-term goals better than long-term performance. Contributing to the levels of selection debate, we encourage researchers to study the implications of the different timescales involved in the eco-evolutionary process.

Geographic variation in female mate-copying in the species complex of a unisexual fish, Poecilia formosa

Summary The Amazon molly, Poecilia formosa, is a gynogenetic, all-female fish. Its mating system relies on heterospecific matings with males of closely related sexual species. In mixed populations, males mate with conspecific sexual females and heterospecific asexual females. Such matings are not isolated dyadic interactions but rather elements of a communication network, because multiple females can observe these interactions. This is the only known case of heterospecific mate-copying and, thus, a system in which the potential for mate-copying could be influenced by the presence of another species. Here we show that mate-copying is exhibited by the sexual species P. mexicana and P. latipinna, and the asexual P. formosa . Both sexual and asexual females copy each other’s mate choice decisions in sympatry, but females from allopatric populations do not show heterospecific mate-copying. Males benefit from heterospecific matings with Amazon mollies because these increase their attractiveness to the conspecific sexual females. In mixed shoals, mate-copying potentially imposes costs as it increases a male’s attractiveness to heterospecific females. We argue that the net-effect of mate-copying is beneficial to males because the relative strength of mate-copying is lower in Amazon mollies. We hypothesize that an added benefit to males lies in the signal value of copulations.

Stable Inheritance of Host Species-Derived Microchromosomes in the Gynogenetic Fish Poecilia formosa

B chromosomes are additional, usually unstable constituents of the genome of many organisms. Their origin, however, is often unclear and their evolutionary relevance is not well understood. They may range from being deleterious to neutral or even beneficial. We have followed the genetic fate of B chromosomes in the asexual, all-female fish Poecilia formosa over eight generations. In this species, B chromosomes come in the form of one to three tiny microchromosomes derived from males of the host species that serve as sperm donors for this gynogenetic species. All microchromosomes have centromeric heterochromatin but usually only one has a telomere. Such microchromosomes are stably inherited, while the telomereless are prone to be lost in both the soma and germline. In some cases the stable microchromosome carries a functional gene lending support to the hypothesis that the B chromosomes in P. formosa could increase the genetic diversity of the clonal lineage in this ameiotic organism and to some degree counteract the genomic decay that is supposed to be connected with the lack of recombination.

Independent Origins of Allotriploidy in the Fish Genus Poeciliopsis

We examined mitochondrial DNA (mtDNA) sequences and allozymes to assess possible modes of origin, clonal diversity, and evolutionary age in a triploid all-female fish of the genus Poeciliopsis from the state of Sinaloa, Mexico. Analysis of multilocus allozymes revealed that the Rio Mocorito biotype (Poeciliopsis monacha-lucida-viriosa) is trihybrid, carrying haploid genomes from three sexually reproducing species, Poeciliopsis monacha, Poeciliopsis lucida, and Poeciliopsis viriosa. Composite allozyme and mtDNA genotypes identified four clones, all bearing closely related mitochondrial haplotypes originally derived from P. monacha. Apparently these trihybrids arose endemically by addition of a haploid genome from P. viriosa, a local sexual species, to an allodiploid biotype, P. monacha-lucida, also found in the Rio Mocorito. The present analysis clearly revealed that P. monacha-lucida-viriosa arose independently of the two allotriploid biotypes that live in a river to the north (Rio Fuerte). Although the origins of allotriploidy in Poeciliopsis are less constrained phylogenetically and geographically than previously thought, known triploid biotypes all had relatively recent origins, which supports the notion that most asexual lineages are evolutionarily short-lived.

The influence of temperature on sex determination in sockeye salmon Oncorhynchus nerka

Thermolability in phenotypic expression of sex was examined in hime salmon, a land-locked type of sockeye salmon Oncorhynchus nerka, which is a commercially valuable species in cold-water lakes of Japan and is expected to have good potential for aquaculture. Eyed-eggs or alevins of genetically all-female fish were subjected to high temperature. Water temperature was raised from 9 to 18 °C (1) in the middle of the eyed-egg stage, 42 days post-fertilization (dpf); (2) just before starting of hatch, 57 dpf; (3) just after completion of hatch, 78 dpf; or (4) 20 days thereafter, 98 dpf, and kept constant at 18 °C until 63 days after emergence of alevins. Fewer survivals were observed when the temperature was raised early in development. The survival rate in the earliest treatment decreased below 5% at 105 dpf. Observation of gonads at 335 to 456 dpf and 733 dpf revealed that 12 of 14 fish (85.7%) exposed to high temperature on and after 57 dpf exhibited a sex change to phenotypic males. Proportions of sex reversal from the genetical female to the phenotypic male decreased contrary to the survival rates as the temperature treatment started later. Sockeye salmon proved to exhibit thermolability in phenotypic expression of sex, and the temperature-sensitive window was demonstrated to exist in the early life stages on and after 57 dpf, which appears to conform with the exogenous androgen-sensitive window. The new technique through temperature control shown in the present study should contribute to producing all-female populations of this species in a safe, simple, environmental friendly, and acceptable manner.

Unusual triploid males in a microchromosome-carrying clone of the Amazon molly, Poecilia formosa

The Amazon molly, Poecilia formosa, is an all-female fish of hybrid origin which reproduces by gynogenesis, i.e. it depends on sperm of males of closely related species to trigger parthenogenetic development of the embryo. Therefore the offspring is clonal and identical to the mother. In rare cases the exclusion mechanism fails and paternal introgression occurs. This may result either in triploid offspring – if the whole haploid chromosome set of the sperm fuses with the diploid egg nucleus – or in siblings with microchromosomes – if only subgenomic amounts of paternal DNA are included. In one of our diploid, microchromosome-carrying laboratory stocks we observed eight triploid individuals which all developed into males. We investigated the mitotic and meiotic chromosomes, the synaptonemal complex (SC), and sperm production of these males, and compared them to males of the gonochoristic parental species (P. latipinna and P. mexicana) and their hybrids. This comparison revealed that P. formosa males are functional males with reduced effective fertility. They show a deviation from the typical 23 bivalents in the synaptonemal complexes as well as in diakinesis due to the triploid state. They produced offspring but only with gynogenetic Amazon molly females. This shows that the probably aneuploid sperm from P. formosa males can trigger parthenogenetic development of unreduced eggs.