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19 pages, 1080 KiB  
Article
Microplastic Bioaccumulation and Oxidative Stress in Key Species of the Bulgarian Black Sea: Ecosystem Risk Early Warning
by Albena Alexandrova, Svetlana Mihova, Elina Tsvetanova, Madlena Andreeva, Georgi Pramatarov, Georgi Petrov, Nesho Chipev, Valentina Doncheva, Kremena Stefanova, Maria Grandova, Hristiyana Stamatova, Elitsa Hineva, Dimitar Dimitrov, Violin Raykov and Petya Ivanova
Microplastics 2025, 4(3), 50; https://doi.org/10.3390/microplastics4030050 - 6 Aug 2025
Abstract
Plastic pollution in marine environments poses a new global threat. Microplastics (MPs) can bioaccumulate in marine organisms, leading to oxidative stress (OS). This study investigates MP accumulation and associated OS responses in six invertebrate species (Bivalvia, Gastropoda, and Malacostraca) and three key fish [...] Read more.
Plastic pollution in marine environments poses a new global threat. Microplastics (MPs) can bioaccumulate in marine organisms, leading to oxidative stress (OS). This study investigates MP accumulation and associated OS responses in six invertebrate species (Bivalvia, Gastropoda, and Malacostraca) and three key fish species of the Bulgarian Black Sea ecosystems. The target hydrobionts were collected from nine representative coastal habitats of the northern and southern aquatory. MPs were quantified microscopically, and OS biomarkers (lipid peroxidation, glutathione, and antioxidant enzymes) were analyzed spectrometrically in fish liver and gills and invertebrate soft tissues (STs). The specific OS (SOS) index was calculated as a composite indicator of the ecological impact, incl. MP effects. The results revealed species-specific MP bioaccumulation, with the highest concentrations in Palaemon adspersus, Rathke (1837) (0.99 ± 1.09 particles/g ST) and the least abundance in Bittium reticulatum (da Costa, 1778) (0.0033 ± 0.0025 particles/g ST). In Sprattus sprattus (Linnaeus, 1758), the highest accumulation of MPs was present (2.01 ± 2.56 particles/g muscle). The correlation analyses demonstrated a significant association between MP counts and catalase activity in all examined species. The SOS index varied among species, reflecting different stress responses, and this indicated that OS levels were linked to ecological conditions of the habitat and the species-specific antioxidant defense potential to overcome multiple stressors. These findings confirmed the importance of environmental conditions, including MP pollution and the evolutionarily developed capacity of marine organisms to tolerate and adapt to environmental stress. This study emphasizes the need for novel approaches in monitoring MPs and OS to better assess potential ecological risks. Full article
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13 pages, 745 KiB  
Article
Optimizing Selenium Polysaccharide Supplementation: Impacts on Growth, Oxidative Stress, and Tissue Selenium in Juvenile Large Yellow Croaker (Larimichthys crocea)
by Jinxing Xiao, Zhoudi Miao, Shiliang Dong, Kaiyang Wang, Fan Zhou and Zilong Li
Animals 2025, 15(15), 2292; https://doi.org/10.3390/ani15152292 - 6 Aug 2025
Abstract
Selenium (Se) is an essential trace element critical for animal growth and immune function. This study investigated the dietary selenium requirement of juvenile large yellow croaker (Larimichthys crocea) through an 8-week feeding trial. Five experimental diets were formulated by supplementing a [...] Read more.
Selenium (Se) is an essential trace element critical for animal growth and immune function. This study investigated the dietary selenium requirement of juvenile large yellow croaker (Larimichthys crocea) through an 8-week feeding trial. Five experimental diets were formulated by supplementing a basal diet with selenium polysaccharides (Se-PS) at 0, 20, 30, 40, and 50 mg/kg, resulting in analyzed Se concentrations of 0.35, 0.54, 0.71, 0.93, and 1.11 mg/kg, respectively. The results demonstrated that growth performance and feed efficiency improved with increasing dietary selenium, peaking at 0.93 mg/kg before declining at higher levels. Antioxidant enzyme activities—superoxide dismutase (SOD) and catalase (CAT)—in serum and liver tissues exhibited a dose-dependent increase, reaching maximal levels at 1.11 mg/kg. Conversely, malondialdehyde (MDA), a marker of oxidative stress, progressively decreased in both serum and liver, attaining its lowest concentration at 1.11 mg/kg, though this did not differ significantly from the 0.93 mg/kg group (p = 0.056). Tissue selenium accumulation was highest at these optimal dietary levels. Based on the growth performance, oxidative stress response, and tissue selenium retention, the recommended dietary selenium requirement for juvenile large yellow croaker is 0.93 mg/kg. These findings highlight the importance of optimal Se supplementation in aquafeeds to enhance growth and physiological health in farmed fish. Full article
(This article belongs to the Section Aquatic Animals)
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14 pages, 2957 KiB  
Article
Histochemical Study of Enzyme Activity in the Digestive Tract of the Small-Spotted Catshark (Scyliorhinus canicula) and the Smooth-Hound (Mustelus mustelus)
by Lucija Devčić, Ivan Vlahek, Magdalena Palić, Valerija Benko, Siniša Faraguna, Marin Lovrić, Damir Valić and Snježana Kužir
Fishes 2025, 10(8), 386; https://doi.org/10.3390/fishes10080386 - 6 Aug 2025
Abstract
The small-spotted catshark and the smooth-hound are cartilaginous, carnivorous fish with similar depth ranges in their habitats. These two species are among the most abundant elasmobranchs in the Adriatic Sea and are frequently caught by local fishermen using longline fishing. Despite their ecological [...] Read more.
The small-spotted catshark and the smooth-hound are cartilaginous, carnivorous fish with similar depth ranges in their habitats. These two species are among the most abundant elasmobranchs in the Adriatic Sea and are frequently caught by local fishermen using longline fishing. Despite their ecological similarities, little is known about the physiological differences in their digestive processes. The study of enzymatic digestion in these ecologically relevant species helps to fill the knowledge gap in the understanding of nutrient processing in cartilaginous fish. Therefore, the aim of this study was to determine, measure and compare the enzymatic activity of alkaline phosphatase, acid phosphatase, non-specific esterase and aminopeptidase. Fish were caught in the central part of the Adriatic Sea between 2021 and 2023. A total of 60 adult individuals were analyzed, with samples taken from six parts of the digestive tract. Histochemical analysis of 1440 slides revealed clear differences in enzyme activity between the two species. In the small-spotted catshark, cellular protein degradation was most pronounced in esophagus, posterior stomach and rectum, whereas in the smooth-hound, it was concentrated in posterior stomach and spiral intestine. Cellular digestion of lipids in the small-spotted catshark appears to occur primarily in the stomach. The results of this study provide new insights into the distribution of cellular digestive enzymes in cartilaginous fish and emphasize the importance of studying the entire digestive tract as an integrated system rather than focusing on individual parts. This study fills an important knowledge gap and contributes to a deeper understanding of digestive physiology, which in turn has implications for species conservation and biological variability. Full article
(This article belongs to the Section Physiology and Biochemistry)
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12 pages, 2254 KiB  
Article
Evolution of the Jawed Vertebrate (Gnathostomata) Stomach Through Gene Repertoire Loss: Findings from Agastric Species
by Jackson Dann and Frank Grützner
J. Dev. Biol. 2025, 13(3), 27; https://doi.org/10.3390/jdb13030027 - 5 Aug 2025
Abstract
The stomach has been a highly conserved organ throughout vertebrate evolution; however, there are now over 20 lineages composed of monotremes, lungfish and teleost fish displaying a secondary loss of stomach function and morphology. This “agastric phenotype” has evolved convergently and is typified [...] Read more.
The stomach has been a highly conserved organ throughout vertebrate evolution; however, there are now over 20 lineages composed of monotremes, lungfish and teleost fish displaying a secondary loss of stomach function and morphology. This “agastric phenotype” has evolved convergently and is typified by a loss of gastric glands and gastric acid secretion and a near-to-complete loss of storage capacity of the stomach. All agastric species have lost the genes for gastric enzymes (Pga and Pgc) and proton pump subunits (Atp4a and Atp4b), and gastrin (Gast) has been lost in monotremes. As a key gastric hormone, the conservation of gastrin has not yet been investigated in the lungfish or agastric teleosts, and it is unclear how the loss of gastrin affects the evolution and selection of the native receptor (Cckbr), gastrin-releasing peptide (Grp) and gastrin-releasing peptide receptor (Grpr) in vertebrates. Furthermore, there are still many genes implicated in gastric development and function which have yet to be associated with the agastric phenotype. We analysed the evolution, selection and conservation of the gastrin pathway and a novel gastric gene repertoire (Gkn1, Gkn2, Tff1, Tff2, Vsig1 and Anxa10) to determine the correlation with the agastric phenotype. We found that the loss of gastrin or its associated genes does not correlate with the agastric phenotype, and their conservation is due to multiple pleiotropic roles throughout vertebrate evolution. We found a loss of the gastric gene repertoire in the agastric phenotype, except in the echidna, which retained several genes (Gkn1, Tff2 and Vsig1). Our findings suggest that the gastrin physiological pathway evolved differently in pleiotropic roles throughout vertebrate evolution and support the convergent evolution of the agastric phenotype through shared independent gene-loss events. Full article
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20 pages, 450 KiB  
Article
Four Organic Protein Source Alternatives to Fish Meal for Pacific White Shrimp (Penaeus vannamei) Feeding
by Yosu Candela-Maldonado, Imane Megder, Eslam Tefal, David S. Peñaranda, Silvia Martínez-Llorens, Ana Tomás-Vidal, Miguel Jover-Cerdá and Ignacio Jauralde
Fishes 2025, 10(8), 384; https://doi.org/10.3390/fishes10080384 - 5 Aug 2025
Abstract
The use of eco-organic ingredients as a source of protein in aquaculture diets needs important attention due to the growing demand for organic seafood products. The present study evaluated the effects of fish meal substitution by different organic ingredients on the growth, body [...] Read more.
The use of eco-organic ingredients as a source of protein in aquaculture diets needs important attention due to the growing demand for organic seafood products. The present study evaluated the effects of fish meal substitution by different organic ingredients on the growth, body composition, retention efficiency, enzyme activity, and nutrient digestibility of white shrimp Penaeus vannamei. The four dietary formulations tested were formulated with organic ingredients and the fish meal was replaced by the following organic protein meals: Iberian pig viscera meal (PIG), trout by-product meal (TRO), insect meal (FLY), and organic vegetable meal (WHT), in addition to a control diet (CON) that included 15% fish meal. A growth trial was carried out for 83 days, raising 1 g shrimp to commercial size (20 g). Shrimp were stocked at 167 shrimp/m3 (15 individuals per 90 L tank). The results showed that the growth obtained by shrimp fed with TRO (19.27 g) and PIG (19.35 g) were similar in weight gain to the control diet (20.76 g), while FLY (16.04 g) and WHT (16.73 g) meals resulted in a significant lower final weight. The FLY diet showed significantly lower protein digestibility (68.89%) compared to the CON, PIG, TRO, and WHT diets, and significantly higher trypsin activity (0.17 mU/g) compared to shrimp fed with the PIG, TRO, and WHT diets. Shrimp fed with WHT have a significantly lower body weight percentage of protein (19.69%) than shrimp fed with the WHT and TRO diets, and some significant differences in dietary aminoacidic levels affecting amino acid body composition. These results indicate that Iberian pig viscera and trout by-product meal can successfully replace fish meal in Pacific white shrimp aquaculture. Full article
(This article belongs to the Special Issue Advances in Aquaculture Feed Additives)
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19 pages, 7853 KiB  
Article
Pre-Transport Salt Baths Mitigate Physiological Stress and Tissue Damage in Channel Catfish (Ictalurus punctatus) Fingerlings: Evidence from Multi-Biomarker Assessment and Histopathology
by Guowei Huang, Haohua Li, Juguang Wang, Tao Liao, Liang Qiu, Guangquan Xiong, Lan Wang, Chan Bai and Yu Zhang
Animals 2025, 15(15), 2249; https://doi.org/10.3390/ani15152249 - 31 Jul 2025
Viewed by 136
Abstract
Effective transport strategies are critical for the survival and welfare of juvenile Ictalurus punctatus, but the effects of pre-transport salt bath treatments remain uncertain. In this study, we systematically evaluated the effects of pre-transport salt bath acclimation at 0‰ (S1), 1‰ (S2), [...] Read more.
Effective transport strategies are critical for the survival and welfare of juvenile Ictalurus punctatus, but the effects of pre-transport salt bath treatments remain uncertain. In this study, we systematically evaluated the effects of pre-transport salt bath acclimation at 0‰ (S1), 1‰ (S2), 5‰ (S3), and 9‰ (S4) salinity for 30 min on stress resilience and recovery in fingerlings during 12 h of simulated transport and 24 h of recovery. All fish survived, but total ammonia nitrogen (TAN) increased, and pH decreased in all groups, except S3, which showed significantly lower TAN and higher pH (p < 0.05). The S3 and S4 groups showed attenuated increases in serum cortisol and glucose, with S3 exhibiting the fastest return to baseline levels and stable serum sodium and potassium levels. Liver antioxidant enzyme activities in group S3 remained stable, with the lowest malondialdehyde (MDA) accumulation. Integrated biomarker response (IBR) and histological analyses demonstrated that S3 had the lowest systemic stress and tissue damage, whereas S1 and S4 displayed marked cellular disruption. These results indicate that a 5‰ salt bath applied prior to transport may improve water quality, mitigate stress responses, and preserve tissue integrity in juvenile channel catfish. Further studies are needed to confirm these findings in other species and under commercial transport conditions. Full article
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14 pages, 1284 KiB  
Article
Non-Enzymatic Selective Detection of Histamine in Fishery Product Samples on Boron-Doped Diamond Electrodes
by Hiroshi Aoki, Risa Miyazaki and Yasuaki Einaga
Biosensors 2025, 15(8), 489; https://doi.org/10.3390/bios15080489 - 29 Jul 2025
Viewed by 237
Abstract
Histamine sensing that uses enzymatic reactions is the most common form of testing due to its selectivity for histamine. However, enzymes are difficult to store for long periods of time, and the inactivation of enzymes decreases the reliability of the results. In this [...] Read more.
Histamine sensing that uses enzymatic reactions is the most common form of testing due to its selectivity for histamine. However, enzymes are difficult to store for long periods of time, and the inactivation of enzymes decreases the reliability of the results. In this study, we developed a novel, quick, and easily operated histamine sensing technique that takes advantage of the histamine redox reaction and does not require enzyme-based processes. Because the redox potential of histamine is relatively high, we used a boron-doped diamond (BDD) electrode that has a wide potential window. At pH 8.4, which is between the acidity constant of histamine and the isoelectric point of histidine, it was found that an oxygen-terminated BDD surface successfully detected histamine, both selectively and exclusively. Measurements of the sensor’s responses to extracts from fish meat samples that contained histamine at various concentrations revealed that the sensor responds linearly to the histamine concentration, thus allowing it to be used as a calibration curve. The sensor was used to measure histamine in another fish meat sample treated as an unknown sample, and the response was fitted to the calibration curve to perform an inverse estimation. When estimated in this way, the histamine concentration matched the certified value within the range of error. A more detailed examination showed that the sensor response was little affected by the histidine concentration in the sample. The detection limit was 20.9 ppm, and the linear response range was 0–150 ppm. This confirms that this sensing method can be used to measure standard histamine concentrations. Full article
(This article belongs to the Special Issue Advanced Biosensors for Food and Agriculture Safety)
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12 pages, 953 KiB  
Article
Recovery of Male Siamese Fighting Fish (Betta splendens) After Overland Shipping
by Karun Thongprajukaew, Saowalak Malawa, Sukanya Poolthajit, Nutt Nuntapong and Waraporn Hahor
Animals 2025, 15(14), 2156; https://doi.org/10.3390/ani15142156 - 21 Jul 2025
Viewed by 377
Abstract
Ornamental fish shipped by road or rail may spend days in transit without food, leading to a reduction in somatic growth after transportation and during acclimatization. In the present study, a time-series (0, 2, 4, 6, 8, 10, and 12 days) experiment was [...] Read more.
Ornamental fish shipped by road or rail may spend days in transit without food, leading to a reduction in somatic growth after transportation and during acclimatization. In the present study, a time-series (0, 2, 4, 6, 8, 10, and 12 days) experiment was conducted to investigate the growth recovery of male Siamese fighting fish (Betta splendens, 1.56 ± 0.02 g body weight, n = 15 per group) transported by road for two days. Biometric changes, nesting activity, skin pigmentation, digestive enzyme activity, muscle quality, and whole-body composition, were compared across all fish groups. The recovery in growth, as indicated by final body weight, increased with post-transportation time (p < 0.05), causing a significant reversal of weight loss with a proportionally stable condition factor from day 8 until the end of observation (p > 0.05). During this time period, the fish exhibited similar bubble-nest building activity to the control group that was not transported (p > 0.05). Color parameters, digestive enzyme activities, muscle quality, and whole-body composition of fish 8 days after shipping were comparable to the control fish group (p > 0.05). Our findings indicate that an 8-day recovery time is an appropriate protocol for Siamese fighting fish acclimatization following overland shipping. Full article
(This article belongs to the Section Animal Physiology)
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22 pages, 2538 KiB  
Article
Modulation of Digestive Enzyme Activities and Intestinal γ-Proteobacteria in Gilthead Sea Bream Fed High-Fat Diets Supplemented with HIDROX® Olive Oil Extract
by Irene García-Meilán, Sara Balbuena-Pecino, Manel Montblanch, Sara Ramos-Romero, Ramón Fontanillas, Joaquim Gutiérrez, Encarnación Capilla, Isabel Navarro and Ángeles Gallardo
Animals 2025, 15(14), 2102; https://doi.org/10.3390/ani15142102 - 16 Jul 2025
Viewed by 433
Abstract
High-fat diets are commonly used in fish farming due to their protein-sparing effect, contributing to reduced production costs. However, this practice may have adverse effects such as metabolic impairment and inflammation. These problems can be assessed in two ways: by developing functional diets [...] Read more.
High-fat diets are commonly used in fish farming due to their protein-sparing effect, contributing to reduced production costs. However, this practice may have adverse effects such as metabolic impairment and inflammation. These problems can be assessed in two ways: by developing functional diets or using food restriction, which leads to compensatory growth. The present study characterized digestion in gilthead sea bream fed a high-fat diet in the presence (HT) or absence (HF) of an olive oil polyphenol as an additive, hydroxytyrosol, under two different dietary regimes: feeding to satiation (ST) or at a 40% restriction (R). Digestive enzyme activities, specifically trypsin-like activities, were mainly upregulated by dietary treatment (HT). In contrast, restriction effects mainly appeared during digestion in the pyloric caeca, where a significant rise in chymotrypsin-like activities was detected. Moreover, those fish tended to have an increased relative intestinal length compared to those fish fed at a standard ration. Feed restriction enhanced the growth of γ-Proteobacteria in pyloric caeca and proximal intestinal regions, without altering their population in the distal intestine. Overall, it is suggested that hydroxytyrosol inclusion at a standard ration could improve digestion processes in gilthead sea bream fed high-fat diets under healthier conditions than without this additive. Full article
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13 pages, 1242 KiB  
Article
The Investigation of the Effect of Feeds with Probiotic Additives on Growth and Functional State of Juvenile Steelhead Salmo (Salmo gairdneri)
by Elena N. Ponomareva, Marina N. Sorokina, Vadim A. Grigoriev, Mariya S. Mazanko, Vladimir A. Chistyakov and Dmitry V. Rudoy
Fishes 2025, 10(7), 349; https://doi.org/10.3390/fishes10070349 - 14 Jul 2025
Viewed by 216
Abstract
The research on the effect of feed with probiotic additives on the growth and functional state of young steelhead salmon (Salmo gairdneri) is presented in this study. For the first time, target strains selected not only by antagonism to pathogens but [...] Read more.
The research on the effect of feed with probiotic additives on the growth and functional state of young steelhead salmon (Salmo gairdneri) is presented in this study. For the first time, target strains selected not only by antagonism to pathogens but also by their ability to produce lytic enzymes or secondary metabolites with antioxidant activity were used to create probiotic preparations for aquaculture. This study presents findings showing that groups of fish fed probiotic feeds showed an improved growth performance and higher survival rate compared to the control. It was noted that the weights of fish in the first variant and the second variant of the experiment were higher by 8.8% and 6.8%, respectively. This research showed that juvenile steelhead salmon reared with probiotic-supplemented feeds had an improved ability to survive in high salinity and sublethal temperatures. This indicates that probiotics may play a significant role in enhancing the adaptive system of fish. Full article
(This article belongs to the Section Nutrition and Feeding)
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16 pages, 4256 KiB  
Article
Physiological Stress Responses Associated with Microplastic Ingestion in the Benthic Flatfish Bothus podas
by Amanda Cohen-Sánchez, Montserrat Compa, Jessica Lombardo, Maria Magdalena Quetglas-Llabrés, Maria del Mar Ribas-Taberner, Manuel Jiménez-García, Silvia Tejada and Antoni Sureda
Toxics 2025, 13(7), 584; https://doi.org/10.3390/toxics13070584 - 13 Jul 2025
Viewed by 579
Abstract
Bothus podas (wide-eyed flounder) is a benthic flatfish likely exposed to microplastic (MP) pollution. We investigated MP ingestion and associated physiological effects in wild B. podas collected from Mallorca (Balearic Islands), Spain. Markers of oxidative stress, detoxification, and immunity were quantified in intestinal, [...] Read more.
Bothus podas (wide-eyed flounder) is a benthic flatfish likely exposed to microplastic (MP) pollution. We investigated MP ingestion and associated physiological effects in wild B. podas collected from Mallorca (Balearic Islands), Spain. Markers of oxidative stress, detoxification, and immunity were quantified in intestinal, hepatic, and splenic tissues. MPs were observed in the gastrointestinal tracts of 87.5% of the 24 specimens analyzed, with an average of 3.8 ± 0.6 items per fish. Fiber-type MPs predominated in both the gastrointestinal tract (69.6%) and sediment samples (97%). Additionally, micro-Fourier transform infrared spectroscopy analysis confirmed that the majority of ingested MPs were composed of polyethylene, polypropylene, and polyester. Fish were categorized into low (<3 items) and high (≥3 items) MP groups based on the median number of plastic items found in the gastrointestinal tract to assess sublethal impacts. In the gut, high-MP fish exhibited significantly elevated activities of detoxification enzymes: ethoxyresorufin-O-deethylase (phase I) and glutathione s-transferase (phase II), along with increased antioxidant enzyme superoxide dismutase and inflammatory myeloperoxidase. Gut catalase and malondialdehyde (MDA) were not significantly different between groups. In liver tissues, no biomarkers differed significantly with MP exposure. In the spleen, lysozyme and alkaline phosphatase activities were significantly higher in high-MP fish, while splenic MDA remained unchanged. These results indicate that gastrointestinal MP exposure triggers local oxidative stress responses and systemic immune activation in B. podas. Overall, ingestion of environmentally relevant MP levels elicited detoxification and inflammatory responses without significant increases in MDA, an indicator of oxidative damage, highlighting the physiological stress imposed by plastic pollution on benthic fish. Full article
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16 pages, 5425 KiB  
Article
Black Soldier Fly Larvae Meal as a Sustainable Fishmeal Substitute for Juvenile Hybrid Grouper: Impacts on Growth, Immunity, and Gut Health
by Yan Chen, Wenfeng Li, Minyi Zhong, Jun Ma, Bing Chen, Junming Cao, Jiun-Yan Loh and Hai Huang
Fishes 2025, 10(7), 344; https://doi.org/10.3390/fishes10070344 - 11 Jul 2025
Viewed by 317
Abstract
Background: Aquaculture increasingly seeks sustainable alternatives to fishmeal, a key protein source in fish diets. Black Soldier Fly Larvae (BSFL) meal is a promising substitute, but its effects on fish growth, immunity, and gut health need further investigation. This study aimed to evaluate [...] Read more.
Background: Aquaculture increasingly seeks sustainable alternatives to fishmeal, a key protein source in fish diets. Black Soldier Fly Larvae (BSFL) meal is a promising substitute, but its effects on fish growth, immunity, and gut health need further investigation. This study aimed to evaluate the impact of varying BSFL inclusion levels on juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂), a widely farmed species in tropical aquaculture. Methods: Juvenile hybrid grouper were fed diets with four levels of BSFL substitution (0%, 10%, 30%, and 50%) over 56 days. Key metrics such as growth performance, immune function, antioxidant capacity, and gut transcriptome were analyzed. Results: Replacing fish meal with BSFL meal had no significant effect on the survival rate of hybrid grouper (p > 0.05) but significantly affected growth performance, immune function, and antioxidant capacity (p < 0.05). BSFL10 and BSFL30 groups showed good growth and elevated immune enzyme activity, with significantly higher HIS levels (p < 0.05); the Wf of the BSFL10 group was comparable to the control. However, excessive replacement (BSFL50) led to reduced growth (Wf significantly lower, p < 0.05) and increased oxidative stress, as indicated by higher CAT activity (p < 0.05). Transcriptomic analysis revealed upregulation of immune- and metabolism-related genes with increasing BSFL levels, with immune pathways notably activated in the BSFL50 group. Conclusions: BSFL meal is a promising alternative to fishmeal in juvenile hybrid grouper diets, with moderate inclusion (10–30%) being most beneficial. Excessive BSFL substitution (50%) may impair fish health, highlighting the need for careful formulation in aquaculture diets. Full article
(This article belongs to the Section Nutrition and Feeding)
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55 pages, 3773 KiB  
Review
Molecular Mechanisms and Biomarker-Based Early-Warning Indicators of Heavy Metal Toxicity in Marine Fish
by Andra Oros, Valentina Coatu, Nicoleta Damir, Diana Danilov, Elena Ristea and Luminita Lazar
Fishes 2025, 10(7), 339; https://doi.org/10.3390/fishes10070339 - 10 Jul 2025
Viewed by 602
Abstract
Heavy metals are among the most persistent and bioaccumulative pollutants in marine ecosystems, posing significant toxicological threats to fish via complex molecular and cellular disruptions. This review synthesizes current knowledge on the cascade of mechanistic responses in marine fish following HM exposure, which [...] Read more.
Heavy metals are among the most persistent and bioaccumulative pollutants in marine ecosystems, posing significant toxicological threats to fish via complex molecular and cellular disruptions. This review synthesizes current knowledge on the cascade of mechanistic responses in marine fish following HM exposure, which includes oxidative stress, modulation of antioxidant responses, activation of detoxification systems, DNA damage, inflammation, apoptosis, neuroendocrine disruption, and ultimately, cellular energy imbalance. In addition to established pathways, the review highlights recent advances in mechanistic understanding and biomarker development, including cellular stress responses, epigenetic regulation, metal homeostasis mechanisms, and novel molecular indicators. These mechanisms support the development of an integrated biomarker framework that combines classical indicators (e.g., antioxidant enzymes, metallothionein) with next-generation endpoints (e.g., miRNA profiles, gene-level responses of metal transporters or stress chaperones, epigenetic alterations). The interpretation of biomarker responses requires consideration of the exposure context, environmental variables, and physiological status to ensure accurate assessment of sublethal toxicity in field settings. By bridging mechanistic understanding with biomonitoring relevance, this review provides a comprehensive foundation for advancing molecular tools in pollution monitoring and risk assessment. Special emphasis is placed on biomarkers specific to heavy metal exposure, enhancing their diagnostic value relative to general stress indicators. Full article
(This article belongs to the Section Environment and Climate Change)
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20 pages, 7063 KiB  
Article
Toxicity Responses from Tributyltin Chloride on Haarder (Planiliza haematocheila) Livers: Oxidative Stress, Energy Metabolism Dysfunction, and Apoptosis
by Changsheng Zhao, Anning Suo, Dewen Ding and Wencheng Song
Curr. Issues Mol. Biol. 2025, 47(7), 526; https://doi.org/10.3390/cimb47070526 - 8 Jul 2025
Viewed by 285
Abstract
In coastal waters, tributyltin chloride (TBTC), a persistent organic pollutant, is extensively present. It is uncertain, therefore, if exposure to TBTC can harm haarders and how. This study exposed the fish for 60 days in order to investigate the molecular mechanism of haarder [...] Read more.
In coastal waters, tributyltin chloride (TBTC), a persistent organic pollutant, is extensively present. It is uncertain, therefore, if exposure to TBTC can harm haarders and how. This study exposed the fish for 60 days in order to investigate the molecular mechanism of haarder following TBTC poisoning. Our findings demonstrated that growth indices dropped, liver tissue was damaged, and the liver’s total tin concentration rose following TBTC exposure. Furthermore, we discovered that blood reactive oxygen species rose while total blood cell count decreased. As malondialdehyde levels rose, total antioxidant capacity and antioxidant enzyme activity (superoxide dismutase, catalase, and glutathione peroxidase) were markedly reduced. After being exposed to TBTC, liver cells displayed clear signs of apoptosis. Differentially expressed genes were primarily linked to oxidative stress, energy metabolism, and apoptosis, according to the transcriptome study of livers. Overall, the long-term stress of TBTC resulted in the antioxidant system being harmed, as well as serious malfunction of the energy metabolism and apoptotic response. Full article
(This article belongs to the Special Issue Advances in Molecular Biology Methods in Hepatology Research)
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23 pages, 5105 KiB  
Article
Behavioral, Hematological, Histological, Physiological Regulation and Gene Expression in Response to Heat Stress in Amur Minnow (Phoxinus lagowskii)
by Weijie Mu, Jing Wang, Yanyan Zhou, Shibo Feng, Ye Huang and Qianyu Li
Fishes 2025, 10(7), 335; https://doi.org/10.3390/fishes10070335 - 8 Jul 2025
Viewed by 407
Abstract
Rising water temperatures due to climate change pose a significant threat to Phoxinus lagowskii, a cold-water fish that is ecologically vital to the high-latitude regions of China. This study assessed heat stress effects on behavioral, hematological, histological, physiological, and molecular responses in [...] Read more.
Rising water temperatures due to climate change pose a significant threat to Phoxinus lagowskii, a cold-water fish that is ecologically vital to the high-latitude regions of China. This study assessed heat stress effects on behavioral, hematological, histological, physiological, and molecular responses in P. lagowskii. The critical maximum temperature (CTmax) was determined using the loss of equilibrium (LOE) method, with the CTmax reaching 29 °C. Elevated temperatures lead to an increase in the OBR. Fish were subjected to acute heat stress at 28 °C (below CTmax) for 48 h, with samples collected during the 48 h period. RBC, WBC, HGB, and HCT significantly increased during heat stress but decreased 12 h after heat stress. The levels of serum cortisol and blood glucose after heat stress were significantly higher than those in the control group. After heat stress, the height of the ILCM in the gills increased significantly, and the liver exhibited vacuolar degeneration and hypopigmentation. The activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase in the gills initially increased and then decreased over the duration of heat stress. Most enzyme activities (PK, LDH, PFK, and HK) decreased during heat stress, while LPL and HL levels increased, indicating that lipid metabolism was the primary utilization process under heat stress. There was an increase in SOD activity at 12 h, followed by a decrease at 24 h, and an increase in CAT activity under heat stress. Integrated biomarker response (IBR) and principal component analysis (PCA) were employed to synthesize multi-level responses. The IBR values reached their peak at 3 h and 48 h of heat stress. We observed an upregulation of heat shock proteins (Hsp70, Hsp90, and Hsc70) as well as interleukin-10 (IL-10) in response to heat stress. Our findings offer novel insights into the mechanisms underlying the heat stress response in P. lagowskii, thereby enhancing our understanding of the effects of heat stress on cold-water fish. Full article
(This article belongs to the Special Issue Environmental Physiology of Aquatic Animals)
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