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Macroalgae plays a significant role in the formulation of innovative and environmentally sustainable approaches to address food challenges. Specifically, green macroalgae serve as dietary supplements aimed at improving the health, growth, and feeding efficiency of various species of marine and freshwater fishes, as well as mollusks. The effects of Chaetomorpha linum extract (CLE) on growth performance, physiological responses, and disease resistance are studied in Bellamya bengalensis against Aeromonas hydrophila. In this experiment, adult B. bengalensis (4412 ± 165.25 mg) were randomly divided into 15 rectangular glass aquariums (35 snail/aquaria; 45 L capacity) and their basal diet was supplemented with different levels of CLE, including 0 (CLE0), 1 (CLE1), 2 (CLE2), 3 (CLE3), and 4 (CLE4) g/kg for 60 days. The growth performance in the CLE3 dietary group was significantly higher that of the CLE0 group, exhibiting both linear and quadratic trends in relation to dietary CLE levels (p < 0.05). The activities of pepsin, amylase, and lipase were found to be highest in CLE3 and lowest in CLE0. Both linear and quadratic responses to dietary CLE levels in digestive enzymes were observed (p < 0.05). The activities of superoxide dismutase and catalase in the hepatopancreas were found to be elevated in snails due to the synergistic effect of the supplemented CLE diet. Among different levels of diet given, CLE2-supplemented snails showed an increase in their enzyme activity (p < 0.05). Interestingly, all the CLE-treated snails expressed elevated levels of mucus lysozyme and mucus protein when compared to control (p < 0.05). Additionally, hepatopancreatic acid phosphatase and alkaline phosphatase activity were elevated in snails consuming CLE3 (p < 0.05). The transcription levels of immune-related genes, including mucin-5ac and cytochrome, were significantly elevated in snails that were fed a diet supplemented with 2–4 g of CLE/kg. Furthermore, the transcription level of the acid phosphatase-like 7 protein gene also increased in snails receiving CLE-supplemented diets. After a 14-day period of infection, snails that consumed a diet supplemented with 3–4 g/kg of CLE exhibited a notable increase in survival rates against virulent A. hydrophila. Based on the above findings, it is suggested that a diet supplemented with 3 g/kg of CLE may enhance growth, antioxidant and immune defense, and disease resistance in the freshwater snail B. bengalensis. Full article

Rice–crab co-culture systems (RC) represent promising sustainable intensification approaches, yet their nitrogen (N) cycling and optimal fertilization strategies remain poorly characterized. In this study, we compared RC with rice monoculture system (RM) across four N gradients (0, 150, 210, and 270 kg N·hm⁻²), assessing N dynamics in field water and N distribution in soil. The results showed that field water ammonium nitrogen (NH₄⁺-N) concentrations increased nonlinearly, showing sharp increases beyond 210 kg N·hm⁻². Notably, crab activity in the RC altered the N transformation and transport processes, leading to a prolonged presence of nitrate nitrogen (NO₃⁻-N) in field water for two additional days after tillering fertilization compared to RM. This indicates a critical window for potential nitrogen loss risk, rather than enhanced retention, 15 days after basal fertilizer application. Compared to RM, RC exhibited enhanced nitrogen retention capacity, with NO₃⁻-N concentrations remaining elevated for an additional two days following tillering fertilization, suggesting a potential critical period for nitrogen loss risk. Post-harvest soil analysis revealed contrasting nitrogen distribution patterns: RC showed enhanced NH₄⁺-N accumulation in surface layers (0–2 cm) with minimal vertical NO₃⁻-N redistribution, while RM exhibited progressive NO₃⁻-N increases in subsurface layers (2–10 cm) with increasing fertilizer rates. The 210 kg N·hm⁻² rate proved optimal for the RC, producing a rice yield 12.08% higher than that of RM and sustaining high crab yields, while avoiding the excessive aqueous N levels seen at higher rates. It is important to note that these findings are based on a single-site, single-growing season field experiment conducted in Panjin, Liaoning Province, and thus the general applicability of the optimal nitrogen rate may require further validation across diverse environments. We conclude that a fertilization rate of 210 kg N·hm⁻² is the optimal strategy for RC, effectively balancing productivity and environmental sustainability. This finding provides a clear, quantitative guideline for precise N management in integrated aquaculture systems. Full article

The intensification of shrimp aquaculture is crucial for global food security, but poses significant environmental challenges. This review critically assesses the strengths and bottlenecks of two main treatment paradigms: in situ systems, chiefly biofloc technology (BFT), and advanced ex situ systems, such as recirculating aquaculture systems (RASs), constructed wetlands (CWs), and membrane bioreactors (MBRs). Although BFT enables nutrient recycling, it suffers from nitrate accumulation and a high energy demand. Likewise, ex situ technologies can achieve a high treatment efficiency, but contend with high costs, large footprints, or membrane fouling. In this review, we propose the strategic integration of microalgae, representing a universal and synergistic solution for overcoming these disparate bottlenecks. We dissect how a microalgal co-culture can simultaneously remove nitrate and reduce the aeration costs in BFT systems. Furthermore, we explore how microalgae-based units can serve as efficient polishing steps for RASs, enhance the performance of CWs, and mitigate fouling in MBRs. This review delves into the fundamental mechanisms of the microalgal–bacterial symbiosis that underpins these enhancements. Finally, we highlight the valorization of the resulting algal biomass as a high-value aquafeed ingredient, which can transform waste management into a value-creation opportunity. This review aims to provide a comprehensive roadmap for developing next-generation, microalgae-enhanced aquaculture systems. Full article

The accumulation of heavy metals in fish tissues is widely recognized as an indicator of aquatic environmental pollution, and the analysis of their content provides a basis for assessing ecological risk and the safety of aquatic food. The European eel (Anguilla anguilla) is a species frequently used as a bioindicator in environmental studies due to its wide geographic distribution, long life cycle, and high capacity for bioaccumulation of heavy metals in various tissues. The aim of this study was to assess the variation in the accumulation of heavy metals, i.e., mercury (Hg), lead (Pb), arsenic (As), and cadmium (Cd), in the tissues (muscle, liver, gonads, and gills) of European eels caught in two locations in Polish inland waters. The obtained results showed significant differences both in the concentration levels of individual elements and in their co-occurrence in the examined tissues. The statistical methods used, including correlation analysis, heat maps, and principal component analysis (PCA), allowed for a comprehensive assessment of the relationships between metals and the identification of factors differentiating the studied populations. The obtained results clearly indicate that fish residing in similar environments for long periods exhibit significant differences in heavy metal content in various fish tissues. Fish obtained from environments with potentially higher levels of heavy metal inputs, such as the Oder River EMU compared with the Vistula River EMU, showed higher levels of heavy metal accumulation in tissues. This study also found that the concentration of heavy metals tested did not exceed the safe standards for human fish consumption. Full article

Biofouling and microbiologically influenced corrosion (MIC) pose profound allied threats, both visible and invisible, across global industrial and societal infrastructures, encompassing both stationary and mobile systems, such as maritime shipping, aquaculture, offshore and onshore energy platforms, desalination and wastewater treatment and distribution systems [...] Full article

The Na⁺/K⁺-ATPase (NKA) gene encodes a critical membrane transporter that maintains cellular ion homeostasis and plays a pivotal role in osmoregulation and salinity adaptation of aquatic organisms. In this study, we identified and validated SNP markers in the NKA gene associated with low-salinity tolerance in Scylla paramamosain. Four candidate SNPs (g.72037G>T, g.72122G>C, g.74293G>T, and g.74433G>T) were screened and genotyped in low-salinity tolerant and intolerant groups. Association analysis revealed that mutant genotypes at all four loci were significantly enriched in the tolerant group (p < 0.05), with odds ratios (OR) > 1. The tolerant group exhibited higher genetic diversity parameters than the intolerant group. Haplotype analysis showed the GGGG haplotype was dominant in the intolerant group, whereas the other haplotypes were mainly enriched in the tolerant group. The NKA expression in the mutant genotypes was significantly higher than that in the wild genotypes by qRT-PCR. For tolerant individuals, the fast-growing group exhibited higher mutation frequencies than the slow-growing group. Multi-locus analysis achieved substantially more discrimination accuracy than single-locus analysis. These findings demonstrated that these SNPs could be candidate molecular markers for breeding programs in S. paramamosain in low-salinity environments, helping to identify individuals with enhanced salinity tolerance and supporting sustainable aquaculture practices. Full article

The intensification of aquaculture practices has been accompanied by an increased incidence of bacterial diseases, leading to a greater reliance on antibiotics for disease control. Consequently, the widespread and often indiscriminate use of these compounds has contributed to the emergence and dissemination of antibiotic-resistant bacteria within aquaculture systems, posing a serious threat to animal health, environmental sustainability, and public health. In this regard, research efforts have focused on developing alternative strategies to reduce antibiotic use. Natural compounds have gained particular attention due to their well-documented antimicrobial and antibiofilm activities. In this context, the combined application of antibiotics and natural compounds has emerged as a promising approach to enhance antimicrobial efficacy while potentially mitigating the development of resistance. This review synthesizes the current knowledge on antibiotic resistance in aquaculture, highlights the role of biofilm formation as a key resistance mechanism, and critically examines the potential of antibiotic–natural compound combinations against major aquaculture pathogens, with particular emphasis on bacterial growth inhibition, biofilm disruption, and virulence attenuation. Collectively, the evidence discussed underscores the potential of synergistic strategies as a sustainable tool for improving disease management in aquaculture while supporting efforts to limit antibiotic resistance. Full article

Several studies have demonstrated that methionine supplementation in fish diets enhances immune status, inflammatory response, and resistance to bacterial infections by modulating for DNA methylation, aminopropylation, and transsulfuration pathways. However, the immunomodulatory effects of methionine in viral infections remain unexplored. This study aimed to evaluate the effect of methionine supplementation on immune modulation and resistance to the viral haemorrhagic septicaemia virus (VHSV) in rainbow trout (Oncorhynchus mykiss). Two diets were formulated and fed to juvenile rainbow trout for four weeks: a control diet (CTRL) with all nutritional requirements, including the amino acid profile required for the species, and a methionine-supplemented diet (MET), containing twice the normal requirement of DL-methionine. After feeding, fish were bath-infected with VHSV, while control fish were exposed to a virus-free bath. Samples were collected at 0 (after feeding trial), 24, 72, and 120 h post-infection for the haematological profile, humoral immune response, oxidative stress, viral load, RNAseq, and gene expression analysis. In both diets, results showed a peak in viral activity at 72 h, followed by a reduction in viral load at 120 h, indicating immune recovery. During the peak of infection, leukocytes, thrombocytes, and monocytes migrated to the infection site, while oxidative stress biomarkers (superoxide dismutase glutathione S-transferase, and glutathione redox ratio) suggested a compromised ability to manage cellular imbalance due to intense viral activity. At 120 h, immune recovery and homeostasis were observed due to an increase in the amount of nitric oxide, GSH/GSSG levels, leukocyte replacement, monocyte influx, and a reduction in the viral load. When focusing on the infection peak, gene ontology (GO) analysis showed several exclusively enriched pathways in the skin and gills of MET-fed fish, driven by the upregulation of several key genes. Genes involved in recognition/signalling, inflammatory response, and other genes with direct antiviral activity, such as TLR3, MYD88, TRAF2, NF-κB, STING, IRF3, -7, VIG1, caspases, cathepsins, and TNF, were observed. Notably, VIG1 (viperin), a key antiviral protein, was significantly upregulated in gills, confirming the modulatory role of methionine in inducing its transcription. Viperin, which harbours an S-adenosyl-L-methionine (SAM) radical domain, is directly related to methionine biosynthesis and plays a critical role in the innate immune response to VHSV infection in rainbow trout. In summary, this study suggests that dietary methionine supplementation can enhance a more robust fish immune response to viral infections, with viperin as a crucial mediator. The improved antiviral readiness observed in MET-fed fish underscores the potential of targeted nutritional adjustments to sustain fish health and welfare in aquaculture. Full article

Piscine lactococcosis is an emerging bacterial disease that threatens freshwater and marine aquaculture in the Mediterranean region. This study characterized isolates of Lactococcus garvieae and Lactococcus petauri from farmed fish through molecular identification, genomic typing and antimicrobial susceptibility testing. A total of 39 bacterial strains were analyzed using species-specific real-time PCR assays, multilocus sequence typing and broth microdilution to determine minimum inhibitory concentrations. Results suggest a temporal shift in freshwater systems, where L. garvieae predominated in earlier isolates (mainly ST13, CC4), while L. petauri (ST14, CC14) appears as the dominant species in recent years. In marine fish, only L. garvieae was detected, mainly ST95 (CC95), a lineage previously reported in Europe. Molecular variability was found in both species with lineages capable of infecting livestock and humans. Amoxicillin displayed promising results; florfenicol showed moderate activity, while flumequine exhibited no inhibitory effect. Oxytetracycline and trimethoprim–sulfamethoxazole showed variable results requiring prudent use. These region-specific susceptibility profiles provide updated baseline data to guide empirical antimicrobial therapy while awaiting laboratory confirmation, highlighting the evolution of lactococcosis in aquaculture and emphasizing the need for molecular surveillance, antimicrobial stewardship, and vaccine updates within a One Health framework to mitigate impacts on Mediterranean aquaculture and public health. Full article

Although the clownfish, Amphiprion ocellaris (A. ocellaris), is a popular ornamental marine fish worldwide, the mechanisms underlying color pattern variation remain unclear. Given that the Platinum-type clownfish, nearly entirely white, has high economic value, understanding the biological mechanism that accounts for the difference between orange and white colors in A. ocellaris is crucial. To investigate these coloration differences, we performed RNA sequencing analysis and identified differentially expressed genes (DEGs) by comparing white and orange skin samples from three A. ocellaris individuals. A total of 76 DEGs were detected, including 56 downregulated and 20 upregulated genes. DEG sequences were annotated using Danio rerio and Stegastus partitus as reference species, selecting the best hit based on the lowest E-value. A protein–protein interaction (PPI) network and Gene Ontology biological process terms were additionally analyzed. Several DEGs previously reported to be associated with pigmentation, including hpdb, cldn11b, sfrp5, slc2a9, slc2a11b, si:ch211-256m1.8, fhl2, rab38, and ttc39b were identified. Based on the functions of these DEGs, it is inferred that leucophores and xanthophores contribute to both white and orange coloration by modulating related genes, including slc2a11b and slc2a9. Additionally, sfrp5, sost, and sp7 genes were identified to interact with each other in the PPI analysis, with sfrp5 and sost being associated with the Wnt signaling pathway, which contributes to melanocyte specification and osteoblast differentiation. Based on these findings, we propose sost and sp7 as candidate genes that might provide insights relevant to extreme white pigmentation phenotypes, such as those observed in Platinum-type clownfish. For a clearer understanding, further studies integrating quantitative genetics and functional analyses are required. Full article

Antibiotic feeding in shrimp farming is an optional practice conducted with the aim of preventing and controlling bacterial diseases. However, the administration of antibiotics can disrupt the microbiota of both shrimp and surrounding environment, potentially compromising host health. Given the limited effective antibiotic options in aquaculture, it is crucial to evaluate the effects of florfenicol (FF) on the intestinal health of shrimp and the associated microbial communities. This study first investigated the impact of FF on the intestinal structure of Penaeus vannamei over two feeding durations (5 and 10 days), each followed by a 10-day basal diet recovery period. Simultaneously, variations in microbial communities and antibiotic resistance genes (ARGs) in both the intestine and rearing water were explored. The results showed that intestinal damage was aggravated with the extension of FF duration and gradually recovered after FF withdrawal. Significant changes in microbial composition and β-diversity were observed in both the rearing water and intestine following FF feeding. Extending the FF treatment to 10 days led to a reduced abundance of Rhodobacteraceae and an increased abundance of Flavobacteriaceae and Vibrionaceae in the intestine after 10 days of feeding the basic diet, which may pose a potential risk to shrimp health. Based on correlation analysis of ARGs, microbial communities and pathogenic bacteria, we speculated that rearing water may serve as a reservoir for ARGs dissemination compared to the shrimp intestine. These findings are of great importance for assessing the impact of administration duration under the FF therapeutic dose and highlight the potential risks associated with its overuse in shrimp farming. Full article

Macrobrachium rosenbergii is one of the largest and most economically significant freshwater prawns worldwide. Understanding its population genetic structure is essential for optimizing cross-breeding strategies, conserving germplasm resources, and supporting sustainable aquaculture. However, progress in this area has been hindered by the limited availability of reliable molecular markers. In this study, we developed 20 polymorphic microsatellite primer pairs and applied them to assess the genetic diversity of 11 populations collected from China and Southeast Asia (including Jiangsu, Zhejiang, Taiwan, Myanmar, Bangladesh, Sri Lanka, and Thailand). All loci exhibited high levels of polymorphism. The number of alleles (Na) ranged from 5 to 27, while the mean observed heterozygosity (Ho), expected heterozygosity (He), and polymorphism information content (PIC) were 0.570, 0.720, and 0.686, respectively. The genetic differentiation coefficient (Fst) among populations ranged from 0.017 to 0.289. UPGMA clustering revealed that the Myanmar population formed an independent branch, whereas the remaining ten populations clustered together, indicating relatively close genetic relationships among them. Beyond enriching the currently limited molecular marker resources for M. rosenbergii, this study provides essential baseline data for evaluating genetic diversity in existing populations and establishes a solid molecular foundation for future genetic monitoring and breeding programs. Full article

Feminization is an important biotechnological approach in aquaculture for species in which females exhibit superior growth and higher market value. The long-whiskered catfish (Mystus gulio), a euryhaline species cultivated in both monoculture and co-culture systems, contributes to sustainable aquaculture by grazing on uneaten feed and maintaining pond cleanliness. This study evaluated the effects of dietary 17β-estradiol (E2) at 0, 10, 30, and 60 mg/kg, incorporated into conventional and microencapsulated feeds, on the feminization and early growth of M. gulio larvae. Treatments were administered during the weaning stage for 14 and 21 days under controlled rearing conditions. Results showed that larvae fed microencapsulated feed containing 60 mg/kg E2 achieved the highest specific growth rate (26.91 ± 1.92%/day), feed efficiency (164.76 ± 33.23%), and feminization success (99.73 ± 0.04%). Hormonal assays confirmed elevated estradiol and reduced testosterone levels, consistent with ovarian development observed in histological sections. Gene expression analysis further supported these findings through the significant upregulation of cyp19a, erb1, and erb2 mRNA levels. Overall, this study demonstrates that microencapsulated hormone feeding is an effective and environmentally responsible strategy for achieving monosex female populations in M. gulio, enhancing productivity, reproductive performance, and sustainability in aquaculture systems. Full article

In aquaculture, feed production influences nutrition, performance, water quality, and overall profitability. This study evaluated the effects of three levels of internal fat (IF), resulting from the inclusion of 0%, 2%, or 4% fat in the preconditioner during extrusion, and their interaction with two extruder screw configurations: medium-shear (MS) and high-shear (HS), on kibble physical quality and extrusion parameters. Increasing IF resulted in a quadratic increase in amylose–lipid complexation under the HS configuration (p = 0.030; r2 = 0.9) and a linear reduction (p < 0.001) in specific mechanical energy (SME) with a strong negative Pearson correlation (r −0.9; p = 0.009) in both configurations. Fat inclusion also reduced mass temperature and die pressure (p < 0.05), leading to lower starch gelatinization degree (p < 0.05) from 87.9 ± 0.6% to 83.4 ± 0.3% in MS configuration and 95.6 ± 0.7 to 86.3 ± 0.8% in HS configuration, increased bulk and piece density (p < 0.001), and reduced radial expansion (p < 0.001). These changes decreased floatability (p < 0.05) and water stability, increasing mushiness (p < 0.01). Increased shear partially improved SME transfer, starch cooking, expansion, floatability, and mushiness; however, the negative effects of 4% IF could not be fully mitigated. Overall, higher IF compromised kibble structure, starch gelatinization, and floatability, while screw configuration resulted in only a limited compensatory effect. Full article

The severe decrease in natural sturgeon stocks has led to intensified efforts toward the development of sturgeon aquaculture and the application of reproductive biotechnologies to ensure sustainable production. In cultured male Acipenser ruthenus (sterlet), spermiation must be hormonally induced to obtain high-quality semen; however, reducing hormonal dosages while maintaining or improving sperm quality remains a major challenge. In this study, we investigated the effects of reduced doses of carp pituitary extract (CPE) and luteinizing hormone-releasing hormone agonist (LH-RH) combined with Apilarnil supplementation on spermiation and semen quality in A. ruthenus. Semen volume, pH, sperm concentration, total sperm output, total motility, and spermatozoa velocity were evaluated. Administering a reduced CPE dose (1.1 mg·kg⁻¹) combined with 1 g of Apilarnil significantly increased semen volume, sperm concentration, and total sperm output, as well as improved sperm kinematic parameters (total motility and velocity) compared with the conventional hormonal protocol. Furthermore, treatments using 50% of the standard LH-RH dose supplemented with 1 g or 2 g of Apilarnil resulted in significantly higher sperm motility and velocity than observed in the control group. These results demonstrate that Apilarnil supplementation allows for a substantial reduction in exogenous hormonal doses while enhancing spermiation efficiency and semen quality in sterlet, supporting the spermiation-induction protocol for sturgeon aquaculture. Full article