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<journal-id journal-id-type="publisher-id">Front. Aquac.</journal-id>
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<journal-title>Frontiers in Aquaculture</journal-title>
<abbrev-journal-title abbrev-type="pubmed">Front. Aquac.</abbrev-journal-title>
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<issn pub-type="epub">2813-5334</issn>
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<article-id pub-id-type="doi">10.3389/faquc.2026.1730913</article-id>
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<subject>Review</subject>
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<title-group>
<article-title>From genome to phenome: omics perspectives on fish reproduction</article-title>
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<name><surname>Mushtaq</surname><given-names>Malik Wasim</given-names></name>
<xref ref-type="aff" rid="aff1"><sup>1</sup></xref>
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<name><surname>Rather</surname><given-names>Mohd Ashraf</given-names></name>
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<name><surname>Khan</surname><given-names>Irfan Ahmad</given-names></name>
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<name><surname>Bhat</surname><given-names>Hussna</given-names></name>
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<name><surname>Mir</surname><given-names>Ishfaq Nazir</given-names></name>
<xref ref-type="aff" rid="aff2"><sup>2</sup></xref>
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<name><surname>Bhat</surname><given-names>Irfan Ahmad</given-names></name>
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<aff id="aff1"><label>1</label><institution>Division of Fish Genetics and Biotechnology, Faculty of Fisheries, Sher-e-Kashmir University of Agricultural Sciences and Technology (SKUAST) Srinagar</institution>, <city>Kashmir</city>,&#xa0;<country country="in">India</country></aff>
<aff id="aff2"><label>2</label><institution>Division of Fish Nutrition and Biochemistry, Faculty of Fisheries, Sher-e-Kashmir University of Agricultural Sciences and Technology (SKUAST) Srinagar</institution>, <city>Kashmir</city>,&#xa0;<country country="in">India</country></aff>
<author-notes>
<corresp id="c001"><label>*</label>Correspondence: Malik Wasim Mushtaq, <email xlink:href="mailto:malikhamza13@gmail.com">malikhamza13@gmail.com</email>; Irfan Ahmad Bhat, <email xlink:href="mailto:Bhatirfan13@gmail.com">Bhatirfan13@gmail.com</email></corresp>
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<pub-date publication-format="electronic" date-type="pub" iso-8601-date="2026-02-03">
<day>03</day>
<month>02</month>
<year>2026</year>
</pub-date>
<pub-date publication-format="electronic" date-type="collection">
<year>2026</year>
</pub-date>
<volume>5</volume>
<elocation-id>1730913</elocation-id>
<history>
<date date-type="received">
<day>23</day>
<month>10</month>
<year>2025</year>
</date>
<date date-type="accepted">
<day>16</day>
<month>01</month>
<year>2026</year>
</date>
<date date-type="rev-recd">
<day>06</day>
<month>01</month>
<year>2026</year>
</date>
</history>
<permissions>
<copyright-statement>Copyright &#xa9; 2026 Mushtaq, Rather, Khan, Bhat, Mir and Bhat.</copyright-statement>
<copyright-year>2026</copyright-year>
<copyright-holder>Mushtaq, Rather, Khan, Bhat, Mir and Bhat</copyright-holder>
<license>
<ali:license_ref start_date="2026-02-03">https://creativecommons.org/licenses/by/4.0/</ali:license_ref>
<license-p>This is an open-access article distributed under the terms of the <ext-link ext-link-type="uri" xlink:href="https://creativecommons.org/licenses/by/4.0/">Creative Commons Attribution License (CC BY)</ext-link>. The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.</license-p>
</license>
</permissions>
<abstract>
<p>Fish reproduction, a complex biological process, is regulated by diverse molecular, biochemical, and physiological mechanisms. Advances in omics technologies, including genomics, transcriptomics, metabolomics, and phenomics, have provided every possible way to understand the underlying pathways of reproductive development in fish. In contrast to conventional approaches, omics technologies give us a detailed, molecule-by-molecule view of how an organism&#x2019;s energy, hormones, and metabolism change during its most important reproductive phases. Metabolomics and lipidomic studies in fish have revealed the importance of lipid and amino acid metabolism during embryogenesis and ovarian development, while transcriptomics and proteomics helped to identify key genes and proteins involved in steroidogenic pathway and reproductive development. Nutritional omics on the other hand highlights the role of essential fatty acids, in enhancing overall reproductive performance, so the offspring quality. Moreover, omics is transforming genetic conservation by providing a new toolkit to optimize the freezing process and ensure the long-term survival of genetic resources, securing a future for endangered fish germplasm. Together, these strategies do more than optimize aquaculture like, fortify conservation efforts, build sustainable and resilient futures for fish populations worldwide.</p>
</abstract>
<kwd-group>
<kwd>fish</kwd>
<kwd>genomics</kwd>
<kwd>metabolomics</kwd>
<kwd>reproduction</kwd>
<kwd>transcriptomics</kwd>
</kwd-group>
<funding-group>
<funding-statement>The author(s) declared that financial support was not received for this work and/or its publication.</funding-statement>
</funding-group>
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<custom-meta-group>
<custom-meta>
<meta-name>section-at-acceptance</meta-name>
<meta-value>Disease and Health Management</meta-value>
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</front>
<body>
<sec id="s1" sec-type="intro">
<label>1</label>
<title>Introduction</title>
<p>Suboptimal reproductive performance in fish is a major economic limitation of aquaculture by restricting the production of quality seed and limiting population growth (<xref ref-type="bibr" rid="B55">Honji et&#xa0;al., 2025</xref>). Optimum reproductive success in fish is challenging and usually requires specific interventions. These treatments encompass the delivery of precise hormonal therapy, manipulation of environmental factors like photoperiod, temperature, and salinity, and the use of state-of-the-art reproductive drugs in order to drive processes like gonadal development, ovulation, and gamete viability (<xref ref-type="bibr" rid="B84">Mylonas et&#xa0;al., 2010</xref>). Often, multiple hormonal and environmental approaches are used together, each necessitating a basis in scientific data. New research promotes reproductive technologies through the strategic application of hormone therapies in combination with proper brood stock nutrition to support the production of viable gametes and to establish favorable conditions for spawning or spermiation in many teleost species (<xref ref-type="bibr" rid="B2">Adatto et&#xa0;al., 2022</xref>; <xref ref-type="bibr" rid="B36">Fakriadis et&#xa0;al., 2020</xref>; <xref ref-type="bibr" rid="B28">Corriero et&#xa0;al., 2021</xref>; <xref ref-type="bibr" rid="B53">Hilsdorf et&#xa0;al., 2021</xref>; <xref ref-type="bibr" rid="B51">Hernandez de-Dios et&#xa0;al., 2022</xref>). These advances are essential to optimize both gamete production and quality and thus facilitate the propagation of species of economic value (<xref ref-type="bibr" rid="B88">Naylor et&#xa0;al., 2021</xref>; <xref ref-type="bibr" rid="B14">Bobe and Labb&#xe9;, 2010</xref>; <xref ref-type="bibr" rid="B13">Bobe, 2015</xref>). Reproductive dysfunction is often seen in captive conditions, as reported by commercial producers. Female fish may not complete terminal oocyte maturation, ovulation, or spawning, and males usually produce low-quality and inadequate sperm, leading to decreased growth rates as well as overall production efficiency (<xref ref-type="bibr" rid="B136">Zohar and Mylonas, 2001</xref>; <xref ref-type="bibr" rid="B84">Mylonas et&#xa0;al., 2010</xref>; <xref ref-type="bibr" rid="B135">Zohar et&#xa0;al., 2010</xref>).</p>
<p>Aquaculture, defined as the cultivation of aquatic organisms and plants, represents a rapidly advancing discipline that contributes significantly to food security and livelihood enhancement. The sustainable development of this sector necessitates comprehensive assessments of population dynamics, genetic composition, species identification, and the functional characterization of genes. In this context, the application of genetics and biotechnology provides essential tools for advancing research and innovation (<xref ref-type="bibr" rid="B111">Tripathy et&#xa0;al., 2022</xref>). Omics technologies encompass a suite of high-throughput approaches designed to investigate the structure, function, and interactions of biomolecules, including genes, transcripts, proteins, lipids, and metabolites, within biological systems in an unbiased manner (<xref ref-type="bibr" rid="B56">Horgan and Kenny, 2011</xref>). Following the completion of the Human Genome Project, these approaches rapidly expanded to include genomics, transcriptomics, proteomics, metabolomics, and lipidomics, each providing complementary insights into cellular and molecular processes. In aquaculture, omics-based approaches are increasingly employed for diverse applications. These include deciphering the molecular basis of disease resistance and stress adaptation, developing disease diagnostics and vaccines, identifying and improving resistant strains, authenticating species for food safety and traceability, and improving post-harvest quality and value addition (<xref ref-type="bibr" rid="B120">Westerhoff and Palsson, 2004</xref>).</p>
<p>By integrating data across multiple molecular levels like DNA, RNA, proteins, lipids, and metabolites, omics enables a systems-level understanding of biological complexity. Its utility lies in characterizing global biomolecular states under specific physiological or pathological conditions, identifying key regulatory molecules, and clarifying the mechanisms underlying differential biomolecular responses. The versatility of omics further extends to both fundamental biology and applied research, particularly in disease etiology, biomarker discovery, diagnosis, and prognosis (<xref ref-type="bibr" rid="B61">Kell, 2006</xref>). Future research trajectories are expected to emphasize integrative omics and systems biology approaches, which hold the potential to advance predictive, preventive, and personalized medicine (<xref ref-type="bibr" rid="B35">Evans and Relling, 2004</xref>).</p>
<p>Genomic sequences encode the fundamental blueprint of genetic information; however, the manifestation of performance and production traits is governed by complex regulatory interactions among the genotype, expressed phenotype, and environmental factors (<xref ref-type="bibr" rid="B116">Wang et&#xa0;al., 2009</xref>; <xref ref-type="bibr" rid="B52">Hill and Kirkpatrick, 2010</xref>). These regulatory hierarchies are examined through specialized scientific domains collectively referred to as omics. Within aquaculture systems, the principal omics disciplines encompass genomics, epigenomics, transcriptomics, proteomics, metabolomics, microbiomics, and metagenomics, each providing high-throughput datasets that elucidate specific molecular and cellular components (<xref ref-type="bibr" rid="B78">Mohanty et&#xa0;al., 2019</xref>). Genomics investigates the complete DNA repertoire of an organism, tissue, or cell, facilitating the study of genetic variation, structural organization, and heritability (<xref ref-type="bibr" rid="B83">Mushtaq et&#xa0;al., 2025</xref>). Epigenomics explores chemical and conformational modifications to DNA and RNA that modulate gene regulation, chromatin dynamics, and epigenetic inheritance without altering the nucleotide sequence (<xref ref-type="bibr" rid="B101">Russell, 2010</xref>). Transcriptomics focuses on the global expression profile of RNA transcripts, enabling the quantification of gene activity, alternative splicing patterns, and regulatory mechanisms involving both coding and non-coding RNAs. Proteomics interrogates the entire proteome to characterize protein structures, functions, post-translational modifications, and molecular interactions. Metabolomics assesses the complete spectrum of metabolites, thereby revealing biochemical pathways, cellular fluxes, and metabolic regulation (<xref ref-type="bibr" rid="B30">Davis, 2005</xref>). Microbiomics characterizes the composition, diversity, and functional dynamics of microbial consortia associated with the host and its surrounding environment, whereas metagenomics analyzes the collective genomic content of environmental samples, encompassing genetic material derived from host and non-host organisms alike (<xref ref-type="bibr" rid="B63">Kumar, 2021</xref>). Integration of two or more omics layers constitutes a multi-omics approach, which may also be described as poly-omics, pan-omics, trans-omics, integrative omics, vertical omics, or systems omics, and offers a comprehensive framework for dissecting complex biological processes in aquaculture organisms (<xref ref-type="bibr" rid="B7">Bashura et&#xa0;al., 2021</xref>).</p>
</sec>
<sec id="s2">
<label>2</label>
<title>Genomics in fish reproduction</title>
<p>Genomics deals with the large-scale study of genes and how they are organized within the genome. The genome serves as a long-term storage system for genetic information, but on its own, it does not fully explain how an organism functions. Unlike the proteome, which is dynamic and mirrors active biological processes, the genome remains relatively stable. The genetic information it contains influences an organism&#x2019;s traits by controlling essential cellular activities, mainly through protein synthesis. Genomic research is broadly divided into structural genomics, which examines genome structure, organization, and evolution, and functional genomics, which focuses on patterns of gene expression (<xref ref-type="bibr" rid="B78">Mohanty et&#xa0;al., 2019</xref>). Over time, genomics has shifted from simply sequencing DNA toward understanding gene function. The field now explores how genes are expressed, how they interact in networks, and how these processes connect to biological functions. This expansion has also led to overlaps with proteomics, transcriptomics, and the study of protein&#x2013;protein interactions. Such approaches have opened new opportunities in areas including clinical diagnostics, agricultural and environmental biotechnology, and pharmacogenomics (<xref ref-type="bibr" rid="B76">Misra et&#xa0;al, 2019</xref>). Functional genomics, in particular, aims to study the activity of all genes in a genome at once. This can be examined at the RNA level through transcriptomics or at the protein level through proteomics (<xref ref-type="bibr" rid="B124">Xu et&#xa0;al., 2014</xref>).</p>
<p>A crucial first step in genetic management is setting clear objectives, defining what to manage and how, by integrating ecological, evolutionary and population insights with agreed management priorities. Determining &#x201c;what to manage&#x201d; depends on several factors, including a species&#x2019; physical and behavioral traits, distribution, genetic diversity, ecological role, economic importance, cultural value, population size and structure, and overall extinction risk (<xref ref-type="bibr" rid="B58">Johnson and Koepfli, 2014</xref>). Reproductive technologies now play a vital role in managing both wild and captive populations. Techniques such as artificial insemination and <italic>in vitro</italic> fertilization are increasingly used to promote gene flow among isolated wild populations, between captive and wild groups, and across different conservation programs. These tools also help ensure that genes from individuals unable to breed naturally are still represented in future generations (<xref ref-type="bibr" rid="B93">Pukazhenthi and Wildt, 2004</xref>; <xref ref-type="bibr" rid="B27">Comizzoli et&#xa0;al., 2009</xref>; <xref ref-type="bibr" rid="B121">Wildt et&#xa0;al., 2010</xref>). Several captive-breeding programs have successfully supported or even re-established wild populations. Notable examples include the Puerto Rican parrot (<xref ref-type="bibr" rid="B17">Brock and White, 1992</xref>), California condor (<xref ref-type="bibr" rid="B44">Geyer et&#xa0;al., 1993</xref>), Micronesian kingfisher (<xref ref-type="bibr" rid="B47">Haig et&#xa0;al., 1995</xref>), whooping crane (<xref ref-type="bibr" rid="B59">Jones et&#xa0;al., 2002</xref>), primates such as lion-tailed macaques and bonobos (<xref ref-type="bibr" rid="B81">Morin and Ryder, 1991</xref>; <xref ref-type="bibr" rid="B98">Reinartz and Boese, 1997</xref>), black-footed ferret (<xref ref-type="bibr" rid="B20">Cain et&#xa0;al., 2011</xref>), and the Iberian lynx (<xref ref-type="bibr" rid="B114">Vargas et&#xa0;al., 2008</xref>; <xref ref-type="bibr" rid="B42">Ga&#xf1;&#xe1;n et&#xa0;al., 2010</xref>). The preservation of genetic diversity for future generations is further strengthened through the use of germplasm banks, cryopreserved gametes, and frozen cell lines, supported by ongoing advances in reproductive technologies. Cryo banking is already common in livestock species (<xref ref-type="bibr" rid="B73">Mazur et&#xa0;al., 2008</xref>) and is gradually gaining importance in wildlife conservation as well (<xref ref-type="bibr" rid="B27">Comizzoli et&#xa0;al., 2009</xref>; <xref ref-type="bibr" rid="B107">Swanson et&#xa0;al., 2007</xref>). However, wider application of these methods is often constrained by limited knowledge of the reproductive biology of many species (<xref ref-type="bibr" rid="B6">Andrabi and Maxwell, 2007</xref>).</p>
<p>In fishes, genomics plays an important role in understanding and improving fish reproduction by revealing the genetic basis of reproductive development, maturation, and fertility. Successful reproduction in fish hinges on the precise molecular programming of gonadal development, beginning with the differentiation of bipotential tissue into testes. This process is governed by a complex interaction of genetic cascades and environmental factors. Sex determination can be genotypic (GSD), with master genes like dmy in medaka, amhy in pejerrey, or sdY in salmonids, or environmental (ESD), where factors like elevated temperature promote masculinization often by downregulating ovarian aromatase (cyp19a1a) and upregulating male genes like amh (<xref ref-type="bibr" rid="B72">Matsuda et&#xa0;al., 2002</xref>; <xref ref-type="bibr" rid="B50">Hattori et&#xa0;al., 2012</xref>; <xref ref-type="bibr" rid="B87">Navarro-Mart&#xed;n et&#xa0;al., 2011</xref>).</p>
<p>The differentiation of the testis itself is driven by key transcription factors. Dmrt1 is a pivotal male determinant essential for Sertoli cell specification and spermatogenesis; its disruption leads to testicular dysgenesis and even sex reversal (<xref ref-type="bibr" rid="B118">Webster et&#xa0;al., 2017</xref>; <xref ref-type="bibr" rid="B12">Bhat et&#xa0;al., 2016</xref>). The sox9 genes (duplicated into sox9a and sox9b in teleosts) contribute to Sertoli cell development and testis cord formation, with expression patterns varying between species but often influenced by androgens and temperature (<xref ref-type="bibr" rid="B96">Raghuveer and Senthilkumaran, 2010a</xref>). Anti-M&#xfc;llerian hormone (amh), a member of the TGF-&#x3b2; superfamily, is crucial for testicular differentiation and acts as a negative regulator of spermatogenesis; its expression is upregulated by high temperature and cortisol but typically downregulated by androgens (<xref ref-type="bibr" rid="B91">Pfennig et&#xa0;al., 2015</xref>; <xref ref-type="bibr" rid="B39">Fernandino et&#xa0;al., 2013</xref>).</p>
<p>Steroidogenesis, the production of sex hormones, forms the functional core of the gonad. The pathway from cholesterol to androgens like 11-ketotestosterone (11-KT) involves coordinated expression of genes such as star (cholesterol transport), cyp11a, cyp17, hsd3b, and hsd11b. The final enzyme, aromatase (cyp19a1a), converts androgens to estrogens. This pathway is primarily regulated by the hypothalamic-pituitary-gonadal (HPG) axis. The hypothalamus releases gonadotropin-releasing hormone (GnRH), which stimulates the pituitary to secrete follicle-stimulating hormone (FSH) and luteinizing hormone (LH). These gonadotropins act on testicular somatic cells to drive steroidogenesis and gametogenesis. Kisspeptins (kiss1/kiss2) serve as upstream regulators of GnRH neurons, linking environmental cues like photoperiod to the onset of puberty (<xref ref-type="bibr" rid="B104">Schulz et&#xa0;al., 2010</xref>; <xref ref-type="bibr" rid="B105">Selvaraj et&#xa0;al., 2010</xref>).</p>
<p>Spermatogenesis, the transformation of primordial germ cells (PGCs) into spermatozoa, is the ultimate goal of testicular function. PGCs are marked by genes like vasa and dnd, which are essential for their migration, survival, and early development (<xref ref-type="bibr" rid="B129">Yoshizaki et&#xa0;al., 2002</xref>; <xref ref-type="bibr" rid="B119">Weidinger et&#xa0;al., 2003</xref>). The process is then orchestrated by FSH and locally produced 11-KT, which act through androgen receptors (ar) in Sertoli and germ cells to promote germ cell proliferation and differentiation. Other factors like insulin-like growth factors (igf), activin, and inhibin fine-tune this process by modulating germ cell proliferation and FSH feedback (<xref ref-type="bibr" rid="B104">Schulz et&#xa0;al., 2010</xref>; <xref ref-type="bibr" rid="B77">Miura et&#xa0;al., 1995</xref>).</p>
<p>Critically, the expression of all these genes&#x2014;from sex determiners to steroidogenic enzymes&#x2014;can be modulated by external factors. Hormonal manipulations (e.g., androgen or aromatase inhibitor treatments), environmental changes (temperature, pH), dietary components, and even plant-derived bioactives like eurycomanone can significantly alter transcriptional profiles to either enhance or impair reproductive outcomes (<xref ref-type="bibr" rid="B10">Bhat et&#xa0;al., 2018</xref>; <xref ref-type="bibr" rid="B33">D&#xed;az and Piferrer, 2017</xref>). This molecular understanding provides a roadmap for aquaculture, offering strategies to overcome reproductive dysfunction in captivity by selectively targeting these key genes and pathways to improve gonadal development, steroidogenesis, and ultimately, sustainable gamete production.</p>
<p>Genomics forms the foundation of genome-wide association studies (GWAS) by enabling high-density, genome-wide scanning of single nucleotide polymorphisms (SNPs) to unravel the genetic basis of complex traits such as growth rate and age at sexual maturation in fishes. GWAS and high-resolution QTL mapping in Atlantic salmon (<italic>Salmo salar</italic>) have revealed a well-defined but largely polygenic genetic architecture underlying growth rate and age at sexual maturation. A landmark GWAS by <xref ref-type="bibr" rid="B46">Guti&#xe9;rrez et&#xa0;al. (2015)</xref> identified a major locus on chromosome 25 harboring VGLL3, which explains a substantial proportion of variation in age at maturation, with alleles showing sex-dependent dominance effects on early and late maturation. Adjacent and supporting regions include genes such as SIX6, involved in hypothalamic&#x2013;pituitary axis development, and HIP1, associated with reproductive investment and maturation timing. Growth-related QTLs identified through high-density linkage maps and GWAS overlap with components of the GH&#x2013;IGF axis (<italic>igf1, igf1r, ghr</italic>), as well as metabolic and muscle development genes (<italic>myod, stat5</italic>), indicating tight genetic coupling between somatic growth and reproductive maturation. Similar polygenic patterns for fecundity-related traits have been reported in rainbow trout (<italic>Oncorhynchus mykiss</italic>), where GWAS detected numerous small-effect loci including FSHR, LHR, VIT, ESR1, and BMP15, reinforcing the complexity of female reproductive traits (<xref ref-type="bibr" rid="B29">D&#x2019;Ambrosio et&#xa0;al., 2020</xref>). Collectively, these studies demonstrate that while VGLL3 represents a major-effect maturation gene in Atlantic salmon, most growth and reproductive traits are controlled by many loci of small effect, supporting the use of genomic selection rather than single-marker approaches in salmonid breeding programs (<xref ref-type="bibr" rid="B46">Guti&#xe9;rrez et&#xa0;al., 2015</xref>; <xref ref-type="bibr" rid="B29">D&#x2019;Ambrosio et&#xa0;al., 2020</xref>).</p>
<p>Teleost fishes show remarkable diversity and plasticity in sex determination, with genetic sex determination (GSD) and temperature-dependent sex determination (TSD) operating either independently or in combination across species. Rapid evolution of master sex-determining genes is a characteristic feature of teleosts; the first such gene, DMY, a Y-linked duplicate of DMRT1, was identified in medaka (<italic>Oryzias latipes</italic>) (<xref ref-type="bibr" rid="B72">Matsuda et&#xa0;al., 2002</xref>), whereas closely related species employ alternative regulators such as GsdfY, indicating frequent turnover of sex-determining mechanisms (<xref ref-type="bibr" rid="B85">Myosho et&#xa0;al., 2012</xref>). In salmonids, including rainbow trout and Atlantic salmon, SdY, a duplicated immune-related gene, functions as the master sex-determining gene by interacting with FOXL2 to repress cyp19a1a (aromatase) expression and inhibit ovarian differentiation, thereby promoting testicular development (<xref ref-type="bibr" rid="B126">Yano et&#xa0;al., 2012</xref>, <xref ref-type="bibr" rid="B127">2013</xref>; <xref ref-type="bibr" rid="B8">Bertho et&#xa0;al., 2018</xref>). Increasing evidence demonstrates that epigenetic regulation, particularly DNA methylation, plays a central role in sex determination, differentiation, and temperature-induced sex reversal, with sexually dimorphic methylation patterns often concentrated on sex chromosomes and key pathway genes such as dmrt1 and cyp19a1a showing sex-specific methylation&#x2013;expression relationships. Temperature-driven changes in DNA methylation can bias sex ratios toward masculinization or feminization in a species-specific manner, highlighting the integrated effects of genetics, epigenetics, and environment in teleost sex determination (<xref ref-type="bibr" rid="B74">Metzger and Schulte, 2018</xref>; <xref ref-type="bibr" rid="B125">Yang et&#xa0;al., 2022</xref>).</p>
<p>In recent years, genomics has become a powerful tool in conservation biology, helping to uncover the genetic basis of key life-history and adaptive traits. Although progress has been fastest in model and domestic species where genes related to growth, milk production, and disease resistance have been identified (<xref ref-type="bibr" rid="B37">Fan et&#xa0;al., 2010</xref>). These studies have revealed candidate genes linked to reproduction, development, and behavior (<xref ref-type="bibr" rid="B65">Li et&#xa0;al., 2011</xref>; <xref ref-type="bibr" rid="B75">Miller et&#xa0;al., 2011a</xref>; <xref ref-type="bibr" rid="B103">Sarropoulou and Fernandes, 2011</xref>; <xref ref-type="bibr" rid="B86">Nakamura et&#xa0;al., 2013</xref>). Expanding genomic resources for non-model species remains essential for deepening our understanding of the genetic foundations of key evolutionary traits (<xref ref-type="bibr" rid="B34">Ekblom and Galindo, 2011</xref>).</p>
<sec id="s2_1">
<label>2.1</label>
<title>Genomics and reproductive biology at the cellular level</title>
<p>At the cellular level, reproductive success and, consequently, the fitness and long-term survival of animal populations including fish depend largely on the production of healthy gametes and their ability to create viable offspring (<xref ref-type="bibr" rid="B62">Kime, 1999</xref>). Genomic tools are poised to make some of their most significant contributions to conservation biology by deepening our understanding of the molecular mechanisms underlying reproduction across a wide range of species. Modern high-throughput techniques such as transcriptomics and proteomics are increasingly being used to explore reproductive physiology and uncover the molecular basis of phenomena like reproductive isolation, gamete incompatibility, and related disorders (as illustrated in <xref ref-type="fig" rid="f1"><bold>Figure&#xa0;1</bold></xref>). Comparative genomics, which is inherently iterative, allows researchers to identify candidate genes in one species and then test their functions across evolutionarily distant taxa. For instance, <xref ref-type="bibr" rid="B67">Li et&#xa0;al. (2012)</xref> identified variable markers in 14 candidate genes that are conserved across species ranging from insects to mammals, linking them to male and female fertility outcomes. By integrating data on conserved and divergent traits with genomic features, such as rates of molecular evolution, gene copy number variation, and pseudogenization, scientists can generate testable hypotheses about reproductive mechanisms. When these insights are combined with proteomic datasets, they further help clarify protein&#x2013;protein interactions, including those critical for sperm, egg recognition and fertilization.</p>
<fig id="f1" position="float">
<label>Figure&#xa0;1</label>
<caption>
<p>Genomic methodologies in reproductive biology.</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="faquc-05-1730913-g001.tif">
<alt-text content-type="machine-generated">Semi-circular diagram depicting genomic methodologies advancing reproductive process insights, divided into six color-coded segments: 1. Transcriptomics (blue), characterizing reproductive physiology and mechanisms. 2. Proteomics (light green), elucidating protein interactions. 3. Comparative Genomics (green), testing gene functions across species. 4. Molecular Evolution (yellow), analyzing reproductive phenotypes. 5. Gene Copy Number Variation (orange), identifying fertility-related markers. 6. Pseudogenization (red), revealing non-functional gene influences on fertility.</alt-text>
</graphic></fig>
</sec>
<sec id="s2_2">
<label>2.2</label>
<title>Comparative genomics and adaptive potential</title>
<p>Comparative genomic approaches shed light on the many ways organisms preserve their adaptive potential across different evolutionary contexts. Among vertebrates, the degree of reproductive isolation varies widely: some species diverge rapidly, forming strong postzygotic barriers such as hybrid sterility, while others, like parthenogenetic lizards, maintain hybridization as a recurring mechanism to preserve genetic diversity and reduce inbreeding (<xref ref-type="bibr" rid="B41">Fujita and Moritz, 2009</xref>). Comparative studies of normal and diseased tissues also provide valuable insights, particularly in understanding and managing heritable disorders through improved diagnostic and therapeutic tools. For example, genomic investigations into equine genetic disorders have contributed to better disease management and breeding practices (<xref ref-type="bibr" rid="B18">Brosnahan et&#xa0;al., 2010</xref>). In addition, metagenomic and large-scale ecosystem studies such as the NIH Human Microbiome Project are revealing how microbial communities influence reproductive health, homeostasis, and pathology (<xref ref-type="bibr" rid="B1">Aagaard et&#xa0;al., 2012</xref>). Despite a common vertebrate framework, the embryonic gonad retains dual potential to form either testes or ovaries via distinct pathways, with many non-mammalian vertebrates exhibiting remarkable flexibility beyond chromosomal sex determination. In such species, sex determination is often shaped by environmental and hormonal cues such as temperature, endocrine factors, and chemical exposure reflecting an adaptive response to ecological conditions (<xref ref-type="bibr" rid="B89">Parma and Radi, 2012</xref>; <xref ref-type="bibr" rid="B113">Ungewitter and Yao, 2013</xref>). This plasticity carries important evolutionary consequences. In fishes, for instance, sex ratios are influenced by the combined effects of genetic, environmental, and biochemical factors, resulting in variations in selection pressures and population dynamics across time and space (<xref ref-type="bibr" rid="B92">Piferrer et&#xa0;al., 2012</xref>). Recent genomic technologies are offering unprecedented insights into gametogenic pathways during both embryonic and postnatal development. Transcriptomic analyses of reproductive tissues, when paired with population-level genomic data, have revealed that genes expressed specifically in gonads tend to evolve faster than ovary-biased genes and that reproductive genes in general evolve more rapidly than non-reproductive ones. Interestingly, despite these accelerated evolutionary rates, functional orthologs of reproductive genes maintain broadly similar divergence patterns across vertebrate groups (<xref ref-type="bibr" rid="B45">Grassa and Kulathinal, 2011</xref>). These observations highlight both the dynamic evolutionary nature of reproductive systems and the conserved molecular architecture that supports vertebrate reproduction. Genomic research is also transforming our understanding of epigenetics, the study of how gene activity is regulated without changes to the underlying DNA sequence. Epigenetic modifications involve chemical processes that activate or silence specific genomic regions in a cell- and time-dependent manner. Core mechanisms such as DNA methylation, RNA interference, and histone modification play vital roles in controlling the inheritance and expression of traits with both biological and economic importance (<xref ref-type="bibr" rid="B54">Hong et&#xa0;al., 2011</xref>). Within reproductive genetics, epigenetic regulation is especially important for understanding cellular reprogramming events, including the formation of pluripotent embryonic stem cells and induced pluripotent stem cells, which regain the ability to differentiate into multiple specialized cell types.</p>
</sec>
</sec>
<sec id="s3">
<label>3</label>
<title>Transcriptomics in fish reproduction</title>
<p>The transcriptome represents the complete set of RNA molecules expressed in a cell at a given time, providing a real-time snapshot of gene activity. Transcriptomic approaches especially RNA sequencing (RNA-seq) have become indispensable for studying non-model fish species that lack reference genomes (<xref ref-type="bibr" rid="B43">Garg et&#xa0;al., 2011</xref>). These techniques enable researchers to reconstruct coding regions, detect alternative splicing and post-transcriptional modifications, and quantify gene expression under varying developmental or environmental conditions (<xref ref-type="bibr" rid="B70">Liu et&#xa0;al., 2013</xref>). Unlike earlier microarray-based methods, which required prior genomic information and were limited by hybridization biases and restricted dynamic range (<xref ref-type="bibr" rid="B26">Clark et&#xa0;al., 2002</xref>; <xref ref-type="bibr" rid="B99">Reinartz et&#xa0;al., 2002</xref>; <xref ref-type="bibr" rid="B9">Bertone, 2004</xref>; <xref ref-type="bibr" rid="B49">Harbers and Carninci, 2005</xref>; <xref ref-type="bibr" rid="B94">Qian et&#xa0;al., 2014</xref>), RNA-seq provides far greater accuracy, sensitivity, and resolution for transcriptome profiling (<xref ref-type="bibr" rid="B116">Wang et&#xa0;al., 2009</xref>).</p>
<p>In aquaculture research, transcriptomics is increasingly being used to identify genes associated with key biological processes such as reproduction, growth, stress adaptation, and disease resistance. These insights are accelerating selective breeding programs and contributing to more sustainable production practices. Transcriptome profiling has proven particularly valuable for identifying gene expression patterns and discovering novel molecular markers useful for genetic improvement (<xref ref-type="bibr" rid="B21">Chandhini and Rejish Kumar, 2019</xref>). A notable example comes from research on zebrafish, a well-established model organism in genetics and biotechnology. Extensive transcriptomic analyses in zebrafish have uncovered numerous genes, molecular markers, and expression signatures relevant to developmental and reproductive biology (<xref ref-type="bibr" rid="B115">Vesterlund et&#xa0;al., 2011</xref>). Similarly, transcriptome sequencing of the common caridean shrimp <italic>Crangon crangon</italic> across five developmental stages revealed that nuclear receptors, key biological markers for sex determination, were expressed exclusively in females, alongside RNA interference (RNAi)-related genes. The nuclear receptor superfamily encompasses transcription factors that regulate vital processes such as growth, molting, cell differentiation, and reproduction, while RNAi genes are increasingly being explored as potential antiviral therapeutic targets (<xref ref-type="bibr" rid="B25">Christiaens et&#xa0;al., 2015</xref>). <italic>De novo</italic> transcriptome assemblies are now available for several economically important aquaculture species, including <italic>Lates calcarifer</italic> (<xref ref-type="bibr" rid="B123">Xia and Yue, 2010</xref>), <italic>Oreochromis niloticus</italic> (<xref ref-type="bibr" rid="B132">Zhang et&#xa0;al., 2013</xref>), <italic>Epinephelus coioides</italic> (<xref ref-type="bibr" rid="B57">Huang et&#xa0;al., 2011</xref>), <italic>Ctenopharyngodon idella</italic> (<xref ref-type="bibr" rid="B23">Chen et&#xa0;al., 2012</xref>), <italic>Macrobrachium rosenbergii</italic> (<xref ref-type="bibr" rid="B60">Jung et&#xa0;al., 2011</xref>), <italic>Fenneropenaeus chinensis</italic> (<xref ref-type="bibr" rid="B68">Li et&#xa0;al., 2013a</xref>), <italic>Eriocheir sinensis</italic> (<xref ref-type="bibr" rid="B66">Li et&#xa0;al., 2013b</xref>), and <italic>Crassostrea virginica</italic> (<xref ref-type="bibr" rid="B130">Zhang et&#xa0;al., 2014</xref>) as shown in <xref ref-type="table" rid="T1"><bold>Table&#xa0;1</bold></xref>. Collectively, these studies demonstrate that transcriptome sequencing is a powerful strategy for identifying genes and regulatory pathways associated with desirable traits in aquaculture species particularly those related to immunity, growth, and reproduction. By offering comprehensive insights into gene expression, transcriptomics continues to strengthen the molecular foundation for selective breeding and species improvement programs across aquatic organisms.</p>
<table-wrap id="T1" position="float">
<label>Table&#xa0;1</label>
<caption>
<p>Molecular insights from <italic>de novo</italic> transcriptome research in aquaculture.</p>
</caption>
<table frame="hsides">
<thead>
<tr>
<th valign="top" align="center">Species</th>
<th valign="top" align="center">Purpose of transcriptome study</th>
<th valign="top" align="center">Reference</th>
</tr>
</thead>
<tbody>
<tr>
<td valign="top" align="center"><italic>Lates calcarifer</italic></td>
<td valign="top" align="center">Construction of a <italic>de novo</italic> transcriptome for gene discovery and molecular marker development</td>
<td valign="top" align="center">(<xref ref-type="bibr" rid="B123">Xia and Yue, 2010</xref>)</td>
</tr>
<tr>
<td valign="top" align="center"><italic>Oreochromis niloticus</italic></td>
<td valign="top" align="center">Gene expression profiling for aquaculture traits and adaptation to environmental stress</td>
<td valign="top" align="center">(<xref ref-type="bibr" rid="B132">Zhang et&#xa0;al., 2013</xref>)</td>
</tr>
<tr>
<td valign="top" align="center"><italic>Epinephelus coioides</italic></td>
<td valign="top" align="center">Understanding reproductive biology and immune-related gene expression</td>
<td valign="top" align="center">(<xref ref-type="bibr" rid="B57">Huang et&#xa0;al., 2011</xref>)</td>
</tr>
<tr>
<td valign="top" align="center"><italic>Ctenopharyngodon Idella</italic></td>
<td valign="top" align="center">Study of immune and stress response pathways</td>
<td valign="top" align="center">(<xref ref-type="bibr" rid="B23">Chen et&#xa0;al., 2012</xref>)</td>
</tr>
<tr>
<td valign="top" align="center"><italic>Macrobrachium rosenbergii</italic></td>
<td valign="top" align="center">First large-scale transcriptome for growth and reproduction studies</td>
<td valign="top" align="center">(<xref ref-type="bibr" rid="B60">Jung et&#xa0;al., 2011</xref>)</td>
</tr>
<tr>
<td valign="top" align="center"><italic>Fenneropenaeus chinensis</italic></td>
<td valign="top" align="center">Analysis of immune response and viral defense mechanisms</td>
<td valign="top" align="center">(<xref ref-type="bibr" rid="B68">Li et&#xa0;al., 2013a</xref>)</td>
</tr>
<tr>
<td valign="top" align="center"><italic>Eriocheir sinensis</italic></td>
<td valign="top" align="center">Investigation of developmental and reproductive gene networks</td>
<td valign="top" align="center">(<xref ref-type="bibr" rid="B66">Li et&#xa0;al., 2013b</xref>)</td>
</tr>
<tr>
<td valign="top" align="center"><italic>Crassostrea virginica</italic></td>
<td valign="top" align="center">Study of gene expression under environmental stress and disease challenge</td>
<td valign="top" align="center">(<xref ref-type="bibr" rid="B130">Zhang et&#xa0;al., 2014</xref>)</td>
</tr>
</tbody>
</table>
</table-wrap>
<sec id="s3_1">
<label>3.1</label>
<title>Unraveling reproductive mechanisms through transcriptomics</title>
<p>Reproduction is a fundamental aspect of aquaculture and selective breeding programs, directly determining the success and sustainability of commercial production systems. In controlled hatchery environments, artificial reproduction techniques are routinely applied to enhance spawning efficiency and ensure predictable seed supply. In contrast, in wild populations, accurate species identification becomes critical to prevent unintended hybridization and maintain genetic integrity (<xref ref-type="bibr" rid="B90">Pereiro, 2022</xref>). Many marine species possess wide geographic distributions and complex population structures, which must be carefully considered in both conservation and aquaculture management (<xref ref-type="bibr" rid="B112">Uengwetwanit et&#xa0;al., 2018</xref>). Although artificial reproduction is now commonplace, concerns persist regarding its potential long-term effects on progeny fitness and genetic diversity, particularly under conditions of intensive artificial selection (<xref ref-type="bibr" rid="B125">Yang et&#xa0;al., 2022</xref>). In this context, transcriptomic approaches have emerged as powerful tools to evaluate reproductive performance and elucidate the molecular mechanisms that regulate reproductive physiology and success.</p>
<p>A comprehensive understanding of the genetic loci involved in sex determination is vital for both fisheries management and aquaculture. Fish exhibit remarkable diversity in sex determination systems, ranging from monogenic to polygenic mechanisms located on autosomes or sex chromosomes, often influenced by environmental cues such as temperature, hormones, and other abiotic factors (<xref ref-type="bibr" rid="B32">Devlin and Nagahama, 2002</xref>). Comparative transcriptomic studies between sexes have facilitated the identification of key loci and regulatory pathways associated with sexual differentiation (<xref ref-type="bibr" rid="B24">Chen et&#xa0;al., 2015</xref>; <xref ref-type="bibr" rid="B106">Sun et&#xa0;al., 2013</xref>; <xref ref-type="bibr" rid="B69">Lin et&#xa0;al., 2017</xref>). Analyses of reproductive tissue transcriptomes have also deepened understanding of sex-specific gene expression patterns and the molecular basis of gonadal differentiation (<xref ref-type="bibr" rid="B131">Zhang et&#xa0;al., 2019</xref>; <xref ref-type="bibr" rid="B109">Tao et&#xa0;al., 2018</xref>). Several candidate genes have been consistently identified across fish species (discussed in genomics section) all of which play pivotal roles in sex determination, gametogenesis, and gonadal maturation (<xref ref-type="bibr" rid="B72">Matsuda et&#xa0;al., 2002</xref>; <xref ref-type="bibr" rid="B128">Yokoi et&#xa0;al., 2002</xref>; <xref ref-type="bibr" rid="B126">Yano et&#xa0;al., 2012</xref>; <xref ref-type="bibr" rid="B108">Takehana et&#xa0;al., 2014</xref>; <xref ref-type="bibr" rid="B12">Bhat et&#xa0;al., 2016</xref>, <xref ref-type="bibr" rid="B11">2021</xref>). In the Amur sturgeon (<italic>Acipenser schrenckii</italic>), an economically valuable caviar-producing species, transcriptomic investigations have revealed that long non-coding RNAs (lncRNAs) may modulate gene expression linked to sex-specific traits (<xref ref-type="bibr" rid="B131">Zhang et&#xa0;al., 2019</xref>). Nonetheless, broader transcriptomic analyses across diverse taxa are required to refine our understanding of sex determination and to develop more precise and sustainable breeding strategies. In both wild and cultured fish populations, egg quality remains a decisive factor influencing reproductive success. Considerable variability in egg viability and developmental competence has been documented among species (<xref ref-type="bibr" rid="B22">Chapman et&#xa0;al., 2014</xref>). During early embryogenesis, rapid and synchronized cell divisions occur, relying heavily on maternal mRNA transcripts accumulated during oocyte maturation. These transcripts regulate crucial cellular processes, including cell cycle progression (e.g., cyclins, nucleoplasmin), growth and apoptosis (e.g., insulin-like growth factors, prohibitin), and cytoskeletal organization (e.g., tubulin beta, keratins 8 and 18) (<xref ref-type="bibr" rid="B3">Aegerter et&#xa0;al., 2005</xref>; <xref ref-type="bibr" rid="B15">Bonnet et&#xa0;al., 2007a</xref>, <xref ref-type="bibr" rid="B16">2007b</xref>). In several aquaculture species, such as striped bass, poor egg quality remains a major constraint to production (<xref ref-type="bibr" rid="B22">Chapman et&#xa0;al., 2014</xref>). Comparative transcriptomic studies using model organisms like zebrafish (<italic>Danio rerio</italic>) and medaka (<italic>Oryzias latipes</italic>) have been instrumental in revealing the molecular and developmental pathways governing early reproduction and embryogenesis (<xref ref-type="bibr" rid="B115">Vesterlund et&#xa0;al., 2011</xref>; <xref ref-type="bibr" rid="B95">Qiao et&#xa0;al., 2016</xref>). For instance, gene ontology (GO)-based gene set enrichment analyses (GSEA) using zebrafish annotations have been applied to European glass eels, offering new insights into developmental dimorphism and morphological differentiation (<xref ref-type="bibr" rid="B31">De Meyer et&#xa0;al., 2017</xref>). As genomic and transcriptomic datasets continue to expand, the ability to make robust inferences about fertilization, sex determination, and reproductive physiology in fish will strengthen significantly. These advances will directly support the optimization of spawning management and the enhancement of reproductive efficiency, contributing to more sustainable and productive aquaculture systems worldwide.</p>
</sec>
</sec>
<sec id="s4">
<label>4</label>
<title>Metabolomics in fish reproduction</title>
<p>Metabolomics is the comprehensive study of metabolites, the small molecular substrates, intermediates, and end products of cellular metabolism (<xref ref-type="bibr" rid="B30">Daviss, 2005</xref>). In essence, it captures the unique chemical fingerprints that reflect the physiological state of a cell or organism (<xref ref-type="bibr" rid="B117">Wang et&#xa0;al., 2019</xref>). The metabolome represents the complete set of metabolites within a biological system (cell, tissue, organ, or organism), serving as the final output of diverse biochemical pathways. In aquaculture research, Nuclear Magnetic Resonance (NMR) and Mass Spectrometry (MS) are the two principal analytical platforms used to characterize metabolite profiles. Within this field, two major methodological approaches have gained prominence. The first, metabolic fingerprinting, employs high-throughput analysis across multiple species or experimental conditions to generate predictive models from the full spectrum of detected metabolites. This approach has proven valuable not only in improving aquaculture practices, such as fish health monitoring and farm management, but also in the authentication of fish and feed products (<xref ref-type="bibr" rid="B4">Alfaro and Young, 2018</xref>). The second, metabolic profiling, focuses on the identification and quantification of metabolites through non-targeted analyses of biological systems, thereby revealing global metabolic changes associated with physiological or environmental variations (<xref ref-type="bibr" rid="B100">Roques et&#xa0;al., 2020</xref>). Accurate metabolite identification is critical for elucidating metabolic pathways and understanding how they respond to intrinsic physiological factors (such as growth, reproduction, and stress) or extrinsic environmental stimuli (including diet composition, temperature, and water quality). In aquaculture, metabolomic studies have advanced our knowledge in several key areas, including fish health and welfare, environmental adaptation, and nutritional physiology. In particular, environmental metabolomics has emerged as a powerful subdiscipline, providing insights into how physical, chemical, and biological stressors affect the metabolic homeostasis of aquatic organisms (Bundy et&#xa0;al., 2009). Furthermore, metabolomics has become instrumental in deciphering pathogen-induced metabolic alterations, contributing to improved understanding and management of disease resistance in cultured fish populations. Among the diverse applications of metabolomics, fish nutrition research remains a primary focus. By profiling metabolite changes in response to diet composition and feeding strategies, metabolomics enables a more detailed understanding of nutrient utilization, metabolic efficiency, and the biochemical basis of feed optimization (<xref ref-type="bibr" rid="B111">Tripathy et&#xa0;al., 2022</xref>).</p>
<p>A typical metabolomics workflow involves several sequential stages: sample collection and preparation, analytical measurement, data processing, statistical analysis, and biological interpretation (as illustrated in <xref ref-type="fig" rid="f2"><bold>Figure&#xa0;2</bold></xref>). Muscle and liver tissues are commonly analyzed due to their pivotal roles in metabolic regulation and energy storage. However, alternative biological matrices including plasma, serum, gut contents, feces, mucus, and skin are increasingly employed to investigate pathway-specific responses. To ensure the reliability of metabolomic data, it is crucial to control for potential sources of environmental and biological variability, such as seasonality, tank effects, fish sex, reproductive stage, and water quality parameters, during both experimental design and sampling. Careful standardization at these stages enhances reproducibility and&#xa0;ensures that metabolomic datasets accurately reflect true&#xa0;physiological and environmental effects rather than confounding noise.</p>
<fig id="f2" position="float">
<label>Figure&#xa0;2</label>
<caption>
<p>Workflow of metabolomics approaches in aquaculture research.</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="faquc-05-1730913-g002.tif">
<alt-text content-type="machine-generated">Flowchart illustrating the stages of biological data analysis in a pipeline. Stage 1: Sample Collection and Preparation involves gathering biological samples. Stage 2: Analytical Measurements focuses on testing metabolites. Stage 3: Data Processing entails cleaning data. Stage 4: Statistical Evaluation analyzes data patterns. Stage 5: Biological Interpretation draws conclusions from data. Each stage is connected by colored pipes.</alt-text>
</graphic></fig>
<p>Unlike genes, transcripts, and proteins, metabolites are not directly encoded by the genome. Instead, they encompass a chemically diverse collection of small molecules, such as carbohydrates, amino acids, lipids, nucleotides, and other compound classes, that collectively constitute the metabolome. The metabolome represents the final downstream outcome of gene expression and protein activity, effectively linking molecular processes to cellular function. Because the metabolome integrates inputs from both genetic and environmental factors, its composition tends to fluctuate more rapidly and dramatically than that of the transcriptome or proteome. These variations provide a magnified and highly sensitive snapshot of an organism&#x2019;s physiological state, making metabolomic analysis a powerful approach for detecting subtle biological responses to internal regulation and external stressors (<xref ref-type="bibr" rid="B56">Horgan and Kenny, 2011</xref>).</p>
<sec id="s4_1">
<label>4.1</label>
<title>Metabolomic perspectives on reproductive processes in fish</title>
<p>Metabolomics is fundamentally connected to the phenotypic expression of organisms, offering a powerful framework to understand how they adapt and respond to environmental cues. By examining metabolite composition, this approach provides a precise and dynamic reflection of an organism&#x2019;s physiological state (<xref ref-type="bibr" rid="B40">Fiehn, 2002</xref>). Even subtle variations in genomic or proteomic activity can lead to measurable shifts in metabolite levels, which are often easier to detect and quantify. Compared to genomics and transcriptomics which depend on high-throughput sequencing and extensive reference databases, metabolomics presents a distinct yet complementary methodology. It is at once broader in biological scope and more straightforward in terms of analytical application. Because metabolites are small, chemically diverse molecules that are not constrained by species-specific genetic codes, they serve as versatile and sensitive indicators of biological function (<xref ref-type="bibr" rid="B110">Taylor et&#xa0;al., 2002</xref>). The value of metabolomics becomes especially evident when integrated with other &#x201c;omics&#x201d; approaches. For example, <xref ref-type="bibr" rid="B133">Zhou et&#xa0;al. (2023)</xref> explored energy utilization during embryonic development in Chinese sturgeon (<italic>Acipenser sinensis</italic>) by combining biochemical analyses of embryos at various developmental stages with transcriptomic data. Their results revealed that proteins and lipids act as key energy sources throughout development, with marked variations in amino acid and fatty acid profiles across stages. Notably, lipids were identified as the primary energy substrates during early embryogenesis, highlighting their essential role in fueling rapid cell division and growth. Similarly, studies on ovarian development in Chinese sturgeon have shown that lipid and amino acid metabolism are central to energy supply during reproductive maturation. Analyses of serum metabolomes and ovarian gene expression demonstrated that metabolic activity peaks during stages II and III, when dietary lipids are absorbed and oxidized for energy. Concentrations of histidine, alanine, sarcosine, and multiple fatty acid derivatives reach their highest levels at stage IV, suggesting an increased demand for energy as oocyte maturation progresses. These findings emphasize that lipid and amino acid pathways are fundamental to ovarian development in sturgeons (<xref ref-type="bibr" rid="B134">Zhu et&#xa0;al., 2020</xref>).</p>
<p>Complementary evidence from <xref ref-type="bibr" rid="B71">Leng et&#xa0;al. (2019)</xref> further supports this conclusion, showing that dietary lipid levels significantly influence ovarian development in Chinese sturgeons. Using combined serum metabolomic and gonadal transcriptomic analyses, their study demonstrated that appropriate lipid supplementation enhances reproductive performance and energy metabolism. In another application, <xref ref-type="bibr" rid="B97">Rahimi et&#xa0;al. (2019)</xref> utilized 1H NMR-based metabolomic profiling to examine changes in sperm metabolites of Persian sturgeon following cryopreservation with 2-hydroxypropyl-&#x3b2;-cyclodextrin (H<sub>2</sub>CD) as a cryoprotectant. The inclusion of 10 mM H&amp;#x2082;CD helped preserve several key metabolites such as glucose, guanidinoacetate, O-phosphocholine, and N, N-dimethylglycine while others including lactate, carnitine, and betaine, were less effectively maintained. Although H&amp;#x2082;CD demonstrated partial success in protecting sperm metabolites, its limited efficacy for compounds like creatine phosphate and creatine, which are vital energy substrates, underscores the challenges of maintaining metabolic integrity during cryopreservation. Further integration of lipidomics and metabolomics has provided insights into steroid hormone biosynthesis in sturgeons. <xref ref-type="bibr" rid="B122">Wu et&#xa0;al. (2022)</xref> investigated the role of dietary arachidonic acid [20:4(&#x3c9;-6)] in promoting steroidogenesis in female Chinese sturgeon. Through combined transcriptomic and lipidomic analyses, they identified differentially expressed genes and metabolites across diets containing 0%, 0.5%, 1%, and 2% arachidonic acid. The results revealed that arachidonic acid enhances steroid hormone synthesis by stimulating cholesteryl ester metabolism and upregulating genes involved in steroidogenic pathways. This regulation supports gonadal development and cholesterol metabolism, ultimately improving ovarian maturation and reproductive potential. However, since cholesteryl ester synthesis and hydrolysis are tightly controlled by specific enzymes, the authors emphasized the need for further quantitative analysis of cholesterol esterase and cholesteryl ester hydrolase to validate these mechanisms. <xref ref-type="bibr" rid="B38">Fei et&#xa0;al. (2024)</xref> conducted a study on blotched snakehead (<italic>Channa maculata</italic>) brood stock after fed with a high-protein diet (48%) which significantly enhanced maternal fecundity, egg size, antioxidant capacity, and larval health without affecting fertilization or hatching rates. Integrated metabolomics and transcriptomics revealed that enhanced reproductive performance and egg quality were driven by altered energy and lipid metabolism, particularly arachidonic acid metabolism, steroidogenesis, and TGF-&#x3b2; signaling. Collectively, these findings demonstrate the versatility of metabolomic and lipidomic tools in elucidating the molecular and biochemical foundations of reproduction in aquaculture species. By integrating metabolomic data with transcriptomic and genomic analyses, researchers can better understand how nutritional inputs, environmental stressors, and physiological states shape reproductive performance, providing a strong foundation for optimizing aquaculture management and breeding strategies.</p>
</sec>
</sec>
<sec id="s5">
<label>5</label>
<title>Potential utilization of omics approaches in sustainable aquaculture</title>
<p>The integration of omics technologies into fisheries and aquaculture offers significant benefits for both commercial and conservation-oriented operations, improving product quality, sustainability, and overall competitiveness (<xref ref-type="fig" rid="f3"><bold>Figure&#xa0;3</bold></xref>).</p>
<fig id="f3" position="float">
<label>Figure&#xa0;3</label>
<caption>
<p>Omics technologies in aquaculture.</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="faquc-05-1730913-g003.tif">
<alt-text content-type="machine-generated">Flowchart illustrating the integration of genomic data into aquaculture applications. It starts with information flow from genomic data to integrated analysis and applied outcomes. Key areas include genomics, epigenomics, transcriptomics, proteomics, and metabolomics. These lead to epigenotype and phenome, which contribute to aquaculture applications. Specific applications include trait selection (disease resistance, growth, feed efficiency), breeding programs (genomic prediction), health diagnostics (biomarker discovery), and environmental adaptation (climate resilience).</alt-text>
</graphic></fig>
<p>Omics approaches not only enhance production efficiency but also support the development of farming practices that promote the health and welfare of aquatic organisms, while maintaining environmental sustainability and economic viability. Aquatic species are susceptible to a wide array of pathogens including viruses, bacteria, and parasites many of which remain difficult to detect in the absence of overt clinical signs (<xref ref-type="bibr" rid="B5">Altinok and Kurt, 2003</xref>). Advances in molecular diagnostics, particularly DNA-based tools, now allow for the early detection of pathogens at low concentrations, before disease symptoms manifest.</p>
<p>Current molecular diagnostic strategies in aquaculture primarily include:</p>
<list list-type="order">
<list-item>
<p>Direct sequencing of pathogen DNA, such as ribosomal RNA gene subunits (16S and 23S) for bacterial identification.</p></list-item>
<list-item>
<p>Polymerase chain reaction (PCR), including endpoint PCR (visualized via gel electrophoresis) and real-time PCR using fluorescence-based detection.</p></list-item>
</list>
<p>Nucleic acid microarrays, which enable simultaneous detection of multiple DNA targets or assessment of pathogen gene expression. In parallel, metabolite profiling the measurement of low-molecular-weight compounds and their intermediates provides insight into the physiological responses of organisms to genetic, environmental, and pathological influences. For aquaculture applications, metabolomics must be efficient, reliable, sensitive, and comprehensive. One prominent use is in nutritional studies, where metabolomic and mineral profiling of edible tissues informs dietary value. For instance, <xref ref-type="bibr" rid="B79">Mohanty et&#xa0;al. (2016)</xref> analyzed 35 Indian food fishes and highlighted small indigenous species (SIFs) as rich sources of essential minerals: <italic>Xenentodon cancila</italic> and <italic>Gudusia chapra</italic> for calcium, <italic>Stolephorus waitei</italic> and <italic>X. cancila</italic> for zinc, and <italic>G. chapra</italic> and Penaeus monodon for iron. Such findings underscore the potential of these species to contribute to meeting recommended dietary allowances. The availability of genomic resources for fish has expanded rapidly since the Human Genome Project. In 2014, draft genome sequencing for two key indigenous species, Rohu (<italic>Labeo rohita</italic>) and Magur (<italic>Clarias batrachus</italic>), was initiated by the Rohu and Magur Genome Sequencing Consortium. These reference genomes facilitate whole-genome selection, enabling the identification of genes associated with production traits and supporting selective breeding programs. Additionally, complete mitochondrial genome sequences have been reported for several economically important species, including <italic>Catla catla</italic> (<xref ref-type="bibr" rid="B102">Sahoo et&#xa0;al., 2017</xref>), <italic>Clarias batrachus</italic>, <italic>Pangasius pangasius</italic> (<xref ref-type="bibr" rid="B80">Mohindra et&#xa0;al., 2015</xref>), and Rita rita (<xref ref-type="bibr" rid="B64">Lashari et&#xa0;al., 2015</xref>). These genomic resources enhance the available genetic database for aquaculture, providing essential tools for conservation, genetic improvement, and selective breeding. DNA-based molecular techniques have also become crucial for identifying viral pathogens that threaten aquaculture industries, particularly in cultured finfish and penaeid shrimp. For finfish, diagnostic assays have been developed for pathogens such as infectious hematopoietic necrosis virus, viral <italic>hemorrhagic septicemia</italic> virus, viral nervous necrosis virus (VNNV), and <italic>Renibacterium salmoninarum</italic>. In Japan, PCR screening of striped jack (<italic>Pseudocaranx dentex</italic>) brood stock for VNNV has allowed for the selection of virus-free individuals, reducing the risk of vertical transmission to larvae (<xref ref-type="bibr" rid="B82">Muroga et&#xa0;al., 1998</xref>). Despite these advances, DNA-based approaches have limitations and require careful validation and implementation in aquaculture health management. Beyond genomics, phenomics has emerged as a novel discipline, shifting the focus from single genotype-phenotype relationships to large-scale genome-phenome analyses. This perspective provides a more comprehensive understanding of disease mechanisms and informs strategies for combination therapies (<xref ref-type="bibr" rid="B48">Han et&#xa0;al., 2015</xref>). Traditionally, genotype&#x2013;phenotype interactions were studied via forward genetics (starting from phenotype to gene) or reverse genetics (starting from genotype to phenotype). However, multi-omics approaches have revealed that the complexity of many biological traits and disease phenotypes cannot be fully captured by classical frameworks alone. Instead, integrated genomic, proteomic, and metabolomic datasets are necessary to account for the full spectrum of biological variation, providing a holistic understanding of organismal health, nutrition, and reproductive performance.</p>
</sec>
<sec id="s6">
<label>6</label>
<title>Future perspectives</title>
<p>Future research on fish reproduction should emphasize the integration of multi-omics approaches, including transcriptomics, proteomics, metabolomics, and phenomics, to unravel the intricate molecular and physiological mechanisms underlying gonadal development, gametogenesis, and spawning. The application of advanced molecular diagnostics and metabolite profiling can offer early insights into reproductive health, facilitating precise monitoring of brood stock and timely detection of reproductive disorders. Nutritional omics will be pivotal in designing species-specific diets that optimize reproductive performance and offspring quality, while phenomic approaches can elucidate the complex relationships between genotype, phenotype, and reproductive traits such as fecundity and hormone regulation. Moreover, the integration of omics into cryopreservation and germplasm conservation will support the long-term maintenance of genetic diversity in threatened or commercially valuable species. Together, these strategies promise to advance precision aquaculture, enhancing reproductive efficiency, ensuring genetic sustainability, and supporting the effective conservation and management of fish populations.</p>
</sec>
<sec id="s7" sec-type="conclusions">
<label>7</label>
<title>Conclusion</title>
<p>Omics technologies have emerged as transformative tools for investigating fish reproduction, providing multi-layered insights into the genetic, molecular, and metabolic processes that govern reproductive development. The integrated application of genomics, transcriptomics, proteomics, and metabolomics has enhanced our understanding of how energy substrates, metabolic pathways, and hormonal signals drive embryogenesis, ovarian maturation, and overall reproductive performance. Complementary approaches, such as nutritional and metabolic profiling, have underscored the pivotal role of diet in supporting reproductive efficiency, while cryopreservation and germplasm conservation offer strategies for maintaining genetic diversity. These advances not only improve aquaculture productivity and brood stock management but also support the conservation of endangered fish species. Looking ahead, the continued integration of omics with reproductive biology will be essential for developing sustainable aquaculture strategies that maximize efficiency while safeguarding the long-term resilience of fish populations.</p>
</sec>
</body>
<back>
<sec id="s8" sec-type="author-contributions">
<title>Author contributions</title>
<p>MM: Data curation, Formal Analysis, Conceptualization, Writing &#x2013; original draft. MR: Visualization, Writing &#x2013; review &amp; editing, Methodology. IK: Writing &#x2013; review &amp; editing, Supervision. HB: Writing &#x2013; review &amp; editing, Resources, Data curation. IM:&#xa0;Resources, Data curation, Writing &#x2013; review &amp; editing, Methodology. RB: Writing &#x2013; review &amp; editing, Supervision, Formal Analysis, Visualization, Resources. IB: Methodology, Writing &#x2013; review &amp; editing, Supervision, Conceptualization, Visualization, Validation.</p></sec>
<sec id="s10" sec-type="COI-statement">
<title>Conflict of interest</title>
<p>The author(s) declared that this work was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.</p>
<p>The handling editor SR declared a past co-authorship with the author(s) IB.</p></sec>
<sec id="s11" sec-type="ai-statement">
<title>Generative AI statement</title>
<p>The author(s) declared that generative AI was not used in the creation of this manuscript.</p>
<p>Any alternative text (alt text) provided alongside figures in this article has been generated by Frontiers with the support of artificial intelligence and reasonable efforts have been made to ensure accuracy, including review by the authors wherever possible. If you identify any issues, please contact us.</p></sec>
<sec id="s12" sec-type="disclaimer">
<title>Publisher&#x2019;s note</title>
<p>All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.</p></sec>
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<p><ext-link ext-link-type="uri" xlink:href="https://loop.frontiersin.org/people/865672">Hesham Eed Desouky</ext-link>, Fujian Agriculture and Forestry University, China</p></fn>
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