Effects of Ocimum basilicum essential oil and ginger extract on serum biochemistry, oxidative stress and gill tissue damage of pearl gentian grouper during simulated live transport.

This study explored the effects of the essential oil of Ocimum basilicum (EOOB) and ginger extract (GE) during the transportation of pearl gentian grouper from water quality, serum biochemistry, oxidative stress, meat flavor, and gill tissue morphology. Fish (450 ± 50 g) were allocated to the following 5 treatments: control group (fish transported in water only), 5 mg/LEOOB, 10 mg/LEOOB, 3 mg/LGE, and 6 mg/LGE and transported in insulation boxes (66 × 51 × 37.8 cm) for 72 h. Samples were taken at 0, 12, 36, 60, and 72 h immediately after transport. It was found that 10 mg/LEOOB and 6 mg/LGE could reduce the levels of total ammonia nitrogen (TAN), dissolved oxygen (DO), water pH, serum glucose (GLU), cortisol (COR), liver superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and glutathione peroxidase (GPX), increase the activities of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH), as well as significantly increase the total free amino acid (TFAA) content in muscle compared to the control group (P < 0.05). In addition, by observing the microstructure of gill tissue, it was found that compared with untreated grouper, the morphological damage of gill tissue in EOOB and GE treatment was alleviated. These results indicated that adding appropriate amounts of EOOB and GE to transport water could improve the water quality, relieve stress, and lower energy metabolism of grouper during transport. The results of this research will help to improve the survival rate of grouper after transportation and decrease economic losses to fishery.

Characteristics and molecular properties of crude hemeproteins extracted from Asian seabass gills using an ultrasound-assisted process.

BACKGROUND: The development of a safe and effective iron supplement is important for the treatment of iron-deficient anemia. Therefore, the crude hemeprotein extract (CHPE) from Asian seabass gills was extracted without (CON) and with ultrasound (US)-assisted process, followed by freeze-drying. The resulting freeze-dried crude hemeprotein extract (FDCHPE) powders were determined for trace mineral content, color, secondary structure, protein pattern, size distribution, volatile compounds, and amino acid composition. RESULTS: The extraction yields of CON-FDCHPE and US-FDCHPE were 6.76% and 13.65%, respectively. Highest heme iron (0.485 mg/mL) and non-heme iron (0.023 mg/mL) contents were found when US at 70% amplitude for 10 min (US 70/10) was applied. Both CON-FDCHPE and US-FDCHPE had no heavy metals, but higher iron content (432.8 mg/kg) was found in US-FDCHPE (P < 0.05). Typical red color was observed in CON-FDCHPE and US-FDCHPE with a*-values of 9.72 and 10.60, respectively. Ultrasonication affected protein structure, in which ß-sheet upsurged, whereas random coil, α-helix, and ß-turn were reduced. Protein pattern confirmed that both samples had myoglobin as the major protein. US-FDCHPE also showed a higher abundance of volatile compounds, especially propanal, hexanal, heptanal, and so forth, compared to CON-FDCHPE. Amino acid composition of US-FDCHPE was comparable to Food and Agriculture Organization of the United Nations (FAO) values. CONCLUSION: Overall, FDCHPE extracted using ultrasonication could be safe and effective for fortification in food products as an iron supplement to alleviate iron-deficient anemia. Additionally, gills as leftovers could be better exploited rather than being disposed. © 2023 Society of Chemical Industry.

Roundup® disrupts tissue architecture, attenuates Na+/K+-ATPase expression, and induces protein oxidation/nitration, cellular apoptosis, and antioxidant enzyme expressions in the gills of goldfish, Carassius auratus.

Extensive agricultural activities to feed the growing population are one major driving force behind aquatic pollution. Different types of pesticides are used in farmlands to increase crop production and wash up into water bodies. Glyphosate-based herbicide Roundup® is one of the most used pesticides in the United States; however, its effects on teleost species are still poorly understood. This study focused on the effects of environmentally relevant concentrations of Roundup exposure (low- and high-dose: 0.5 and 5 µg/L for 2-week) on Na+/K+-ATPase (NKA, a biomarker for sodium­potassium ion pump efficacy), cytochrome P450-1A (CYP1A, a monooxygenase enzyme), 2,4-dinitrophenyl protein (DNP, a biomarker for protein oxidation), 3-nitrotyrosine protein (NTP, a biomarker for protein nitration), superoxidase dismutase (SOD, an antioxidant enzyme), catalase (CAT, an antioxidant enzyme) expressions, and cellular apoptosis in the gills of goldfish. Histopathological and in situ TUNEL analyses showed widespread tissue damage, including lamellar fusion, loss of gill architecture, club shape of primary lamellae, mucous formation, and distortion in the epithelium layer, as well as apoptotic nuclei in gills. Immunohistochemical and qRT-PCR analyses provided insights into the expressions of molecular indicators in gills. Fish exposed to Roundup exhibited a significant (P < 0.05) downregulation of NKA expression in gills. Additionally, we observed upregulation of CYP1A, DNP, NTP, SOD, and CAT expressions in the gills of goldfish. Overall, our results suggest that exposure to Roundup causes disruption of gill architecture, induces protein oxidation/nitration and cellular apoptosis, and alters prooxidant-antioxidant homeostasis in tissues, which may lead to reduced fitness and survivability of teleost species.

Aflatoxin B1 damaged structural barrier through Keap1a/Nrf2/ MLCK signaling pathways and immune barrier through NF-κB/ TOR signaling pathways in gill of grass carp (Ctenopharyngodon idella).

Aquafeeds are susceptible to contamination caused by aflatoxin B1 (AFB1). The gill of fish is an important respiratory organ. However, few studies have investigated the effects of dietary AFB1 exposure on gill. This study aimed to discuss the effects of AFB1 on the structural and immune barrier of grass carp gill. Dietary AFB1 increased reactive oxygen species (ROS) levels, protein carbonyl (PC) and malondialdehyde (MDA) contents, which consequently caused oxidative damage. In contrast, dietary AFB1 decreased antioxidant enzymes activities, relative genes expression (except MnSOD) and the contents of glutathione (GSH) (P < 0.05), which are partly regulated by NF-E2-related factor 2 (Nrf2/Keap1a). Moreover, dietary AFB1 caused DNA fragmentation. The relative genes of apoptosis (except Bcl-2, McL-1 and IAP) were significantly upregulated (P < 0.05), and apoptosis was likely upregulated through p38 mitogen-activated protein kinase (p38MAPK). The relative expressions of genes associated with tight junction complexes (TJs) (except ZO-1 and claudin-12) were significantly decreased (P < 0.05), and TJs were likely regulated by myosin light chain kinase (MLCK). Overall, dietary AFB1 disrupted the structural barrier of gill. Furthermore, AFB1 increased gill sensitivity to F. columnare, increased Columnaris disease and decreased the production of antimicrobial substances (P < 0.05) in grass carp gill, and upregulated the expression of genes involved with pro-inflammatory factors (except TNF-α and IL-8) and the pro-inflammatory response partly attributed to the regulation by nuclear factor κB (NF-κB). Meanwhile, the anti-inflammatory factors were downregulated (P < 0.05) in grass carp gill after challenge with F. columnare, which was partly attributed to the target of rapamycin (TOR). These results suggested that AFB1 aggravated the disruption of the immune barrier of grass carp gill after being challenge with F. columnare. Finally, the upper limit of safety of AFB1 for grass carp, based on Columnaris disease, was 31.10 µg/kg diet.

Identification of different physiological functions within the gills and epipodites of the American lobster: Differences in metabolism, transbranchial transport, and mRNA expression.

Transbranchial transport processes are responsible for the homeostatic regulation of most essential physiological functions in aquatic crustaceans. Due to their widespread use as laboratory models, brachyuran crabs are commonly used to predict how other decapod crustaceans respond to environmental stressors including ocean acidification and warming waters. Non-brachyuran species such as the economically-valuable American lobster, Homarus americanus, possess trichobranchiate gills and epipodites that are known to be anatomically distinct from the phyllobranchiate gills of brachyurans; however, studies have yet to define their potential physiological differences. Our results indicate that the pleuro-, arthro-, and podobranch gills of the lobster are functionally homogenous and similar to the respiratory gills of brachyurans as indicated by equivalent rates of H+Eq., CO2, HCO3-, and ammonia transport and mRNA expression of related transporters and enzymes. The epipodites were found to be functionally distinct, being capable of greater individual rates of H+Eq., CO2, and ammonia transport despite mRNA transcript levels of related transporters and enzymes being only a fraction found in the gills. Collectively, mathematical estimates infer that the gills are responsible for 91% of the lobster's branchial HCO3- accumulation whereas the epipodites are responsible for 66% of branchial ammonia excretion suggesting different mechanisms exist in these tissues. Furthermore, the greater metabolic rate and amino acid catabolism in the epipodites suggest that the tissue much of the CO2 and ammonia excreted by this tissue originates intracellularly rather than systemically. These results provide evidence that non-brachyuran species must be carefully compared to brachyuran models.

Accumulation, Chronicity, and Induction of Oxidative Stress Regulating Genes Through Allium cepa L. Functionalized Silver Nanoparticles in Freshwater Common Carp (Cyprinus carpio).

Green evolutionary products such as biologically fabricated nanoparticles (NPs) pose a hazard to aquatic creatures. Herein, biogenic silver nanoparticles (AgNPs) were synthesized by the reaction between ionic silver (AgNO3) and aqueous onion peel extract (Allium cepa L). The synthesized biogenic AgNPs were characterized with UV-Visible spectrophotometer, XRD, FT-IR, and TEM with EDS analysis; then, their toxicity was assessed on common carp fish (Cyprinus carpio) using biomarkers of haematological alterations, oxidative stress, histological changes, differential gene expression patterns, and bioaccumulation. The 96 h lethal toxicity was analysed with various concentrations (2, 4, 6, 8, and 10 mg/l) of biogenic AgNPs. Based on 96 h LC50, sublethal concentrations (1/15th, 1/10th, and 1/5th) were given to C. carpio for 28 days. At the end of experiment, the bioaccumulations of Ag content were accumulated mainly in the gills, followed by the liver and muscle. At an interval of 7 days, the haematological alterations showed significance (p < 0.05) and elevation of antioxidant defence mechanism reveals the toxicity of biogenic synthesized AgNPs. Adverse effects on oxidative stress were probably related to the histopathological damage of its vital organs like gill, liver, and muscle. Finally, the fish treated with biogenic synthesized AgNPs were significantly (p < 0.05) downregulates the oxidative stress genes such as Cu-Zn SOD, CAT, GPx1a, GST-α, CYP1A, and Nrf-2 expression patterns. The present study provides evidence of biogenic synthesized AgNPs influence on the aquatic life through induction of oxidative stress.

Distribution of metals in different environmental compartments and oxidative stress biomarkers in Bryconops caudomaculatus (Osteichthyes: Characiformes) from a bauxite mining area in the Eastern Amazon.

The Eastern Amazon is rich in bauxite ore. The extraction and processing of bauxite lead to the mobilization of Aluminum (Al) and other metals in environmental. We evaluated the metals (Al, Mn, Ba, and Cr) concentration in tissue, water, and sediment associated with antioxidant and oxidative damage responses in Bryconops caudomaculatus. The samplings were done in two hydrological periods (post-rain and post-dry periods) and at three points, located at two rivers: one in the surroundings of the mining area (P1) and other inside the mining area, upstream (P2), and downstream (P3). Defense antioxidant system biomarkers analyzed were total antioxidant capacity (ACAP) and glutathione-S-transferase (GST) activity. As an oxidative damage biomarker, the lipoperoxidation (LPO) was evaluated. Metals concentrations in the water and sediment were higher in the post-rain period compared to post-dry period. The water samples were acidic, with dissolved Al concentrations above the values established by local legislation at all points. In the gills, the metals accumulation was higher in fish from in the surrounding and upstream sites, and in the liver, was higher in fish from downstream site. Fish from the surrounding had increased antioxidant defenses, with higher ACAP in all tissues and higher GST in the gills. Consequently, they had lower levels of LPO. Fish from the mining area had decreased antioxidant defenses, with lower ACAP in all tissues and lower GST in the gills. Consequently, they had higher levels of LPO, indicating oxidative stress. The fish muscle was not responsive to GST and LPO at all sites. We conclude that the oxidative stress observed in the gills and liver of B. caudomaculatus from the area modified by the mining activity reflected the local anthropogenic impact status.

In vivo appraisal of oxidative stress response, cell ultrastructural aberration and accumulation in Juvenile Scylla serrata exposed to uranium.

In vivo studies were performed to evaluate the organ specific tissue accumulation and cellular toxicity of uranium to mud crab Scylla serrata. The specimens were acclimated in natural seawater and the exposure to 50-250 µg/L uranium was investigated up to 60 days. The present study examined the effects of concentration and duration of uranium exposure in the tissue of S. serrata at cellular and subcellular level using scanning electron microscopy and bright field transmission electron microscopy in addition to histological analysis. The results indicated that accumulation of U in S. serrata was organ specific and followed the order gills > hepatopancreas > muscle. The response of key antioxidant enzyme activities such as SOD, GPx and CAT in different organs of crabs indicated oxidative stress due to U in the ambient medium and tissue. At 50 and 100 µg/L of U exposure, individuals were able to acclimate the oxidative stress and withstand the uranium exposure. This acclimation could not be sustained at higher concentrations (250 µg/L), affecting the production of CAT in the tissues. Cellular and subcellular changes were observed in the hemocytes with reduction in their number in consonance with the antioxidant enzymes. Histological aberrations like lamellar disruption of gill, necrosis of hepatopancreas, disruption and rupture of muscle bundles were observed at different concentrations and were severe at higher concentration (250 µg/L). Necrosis was observed in the electron micrographs of tissues shortly after 15 days of exposure. SEM micrograph clearly shows disrupted lamellae, folding of marginal canal and reduction of inter lamellar spaces in the gills of crab exposed to high concentration of uranium. Mitochondrial anomalies are reported for the first time in the present study in addition to the subcellular changes and vacuoles on exposure uranium in the cells of gill and hepatopancreas.

Different toxicity to liver and gill of zebrafish by selenium nanoparticles derived from bio/chemical methods.

With the wide application of selenium nanoparticles (SeNPs) in pharmaceutical fields, the toxicity assessment is of great significance. In this study, zebrafish were selected as model organisms to compare the toxicity of SeNPs derived from biological and chemical methods. The results showed that the size of bio-SeNPs was about 5-fold bigger than chem-SeNPs. When exposed to SeNPs for 96 h, LC50 of bio-SeNPs and chem-SeNPs was 1.668 mg/L and 0.699 mg/L, respectively. Compared with the control, the results showed a significant increase in oxidative toxicity index (P < 0.05), such as glutathione (GSH), superoxide dismutase (SOD) of the liver, and gill in SeNPs-treated group. The neurotoxicity index, such as acetylcholinesterase (AchE) and Na+-K+-ATP enzyme activity, was significantly decreased both in the liver and gill (P < 0.05). It was found that the toxicity of bio-SeNPs to the liver and gill of zebrafish was lower than chem-SeNPs and the toxicity to the liver was higher than gill. In this study, the toxicity of chem-SeNPs and bio-SeNPs to the target organs of zebrafish were systematically evaluated, which provided the basis for the safe application of SeNPs synthesized by different pathways.

Bioremediation of hemotoxic and oxidative stress induced by polyethylene microplastic in Clarias gariepinus using lycopene, citric acid, and chlorella.

Despite extensive research on the toxic effects of microplastics (MPs), there is no obtainable data on the use of phytobioremediation against MPs toxicity in fish. This study aimed to investigate the protective role of lycopene, citric acid, and chlorella against the toxic effects of MPs in African catfish (Clarias gariepinus) using hematology, biochemical, antioxidants, erythron profiles (poikilocytosis and nuclear abnormalities) and the accumulation of MPs in tissues as biomarkers. Five groups of fish received: normal diet (control); MPs (500 mg/kg diet) (Group 2); MPs (500 mg/kg diet) + lycopene (500 mg/kg diet) (Group 3); MPs (500 mg/kg diet) + citric acid (30 g/kg diet) (Group 4); and MPs (500 mg/kg diet) + chlorella (50 g/kg diet) (Group 5) for 15 days. Group 2 had significantly higher amounts of MPs in the stomach, gills, and feces, electrolyte imbalances (HCO3, Fe, Na+, K+, Ca+2, Cl-, and anion gap, hematobiochemical alterations, and decreases in the activities of superoxide dismutase, catalase, total antioxidant capacity, and glutathione S-transferases compared to the control group. Additionally, Group 2 had significant increase in the percentage of poikilocytosis, and nuclear abnormalities in RBC's compared to the control group. The co-treatment of MPs-exposed fish with lycopene, citric acid, and chlorella-supplemented diets ameliorated the hematological, biochemical, and erythron profile alterations, but only slightly enhanced the antioxidant activity. Overall, lycopene, citric acid, and chlorella can be recommended as a feed supplement to improve hematobiochemical alterations and oxidative damage induced by MPs toxicity in the African catfish (C. gariepinus).

Interactive effects of freshwater acidification and selenium pollution on biochemical changes and neurotoxicity in Oreochromis mossambicus.

Effect of selenium and acidification in freshwater environment was assessed solitary but no reports are available on the impacts of both factors act together. In the present study, effects of combined simultaneous exposure to selenium (Se) and low pH were assessed in Mozambique tilapia, Oreochromis mossambicus. Responses were measured based on antioxidant defenses (enzymatic SOD, CAT, GPx and non-enzymatic GSH), biotransformation enzyme (GST), metallothionein levels (MT), oxidative damage (LPO, CP), Na+/K+-ATPase (NKA) activity in gills and liver tissues and neurotoxicity (acetylcholinesterase, AChE) response in brain tissue. Fish were exposed to combined treatment at different pH levels (7.5, control (optimum pH for tilapia growth); 5.5, low pH) and Se concentrations (0, 10, and 100 µg L-1). Toxicity levels of Se were not significantly different under control and low pH indicating that pH did not affect Se toxicity. Levels of GSH and MT were enhanced in Se-exposed fish at both pH. Combined effects of high Se concentration and low pH decreased SOD and CAT activities and increased those of GPx and GST. However, organisms were not able to prevent cellular damage (LPO and CP), indicating a condition of oxidative stress. Furthermore, inhibition of Na+/K+-ATPase activity was showed. Additionally, neurotoxicity effect was observed by inhibition of cholinesterase activity in organisms exposed to Se at both pH conditions. As a result, the combined stress of selenium and freshwater acidification has a slight impact on antioxidant defense mechanisms while significantly inhibiting cholinesterase and Na+/K + -ATPase activity in fish. The mechanisms of freshwater acidification mediating the toxic effects of trace non-metal element on freshwater fish need to investigate further.

Gill oxidative damage caused by acute ammonia stress was reduced through the HIF-1α/NF-κb signaling pathway in golden pompano (Trachinotus ovatus).

This study aimed to investigate the intoxication mechanism of golden pompano (Trachinotus ovatus) exposed to high ammonia levels and the effects on the immune and antioxidant mechanisms of gills. Juvenile golden pompano was exposed to ammonia (total ammonia: 26.9 mg/L) to induce 96 h of ammonia stress, and a 96 h recovery experiment was performed after poisoning. Then, we evaluated hematological parameters, the histological structure and the expression of related genes. In this experiment, continuous exposure to high levels of ammonia led to a significant increase in plasma alkaline phosphatase (ALP), acid phosphatase (ACP) and lactate dehydrogenase (LDH) levels (P < 0.05), and the levels of triiodothyronine (T3) and tetraiodothyronine (T4) were significantly reduced (P < 0.05). Moreover, the expression of antioxidant genes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and inflammatory cytokines such as tumor necrosis factor α (TNF-α) and interleukin 1ß (IL-1ß) increased (P < 0.05). These results indicate that ammonia activates the active osmotic regulatory mechanism of fish gills and participates in defense and immune responses. However, with prolonged exposure to ammonia, the balance of the defense system is disrupted, leading to oxidative damage and inflammation of the gill tissue. This research not only helps elucidate the intoxication mechanism of golden pompano by ammonia at the molecular level but also provides a theoretical basis for further research on detoxification mechanisms.

Adsorptivity of mercury on magnetite nano-particles and their influences on growth, economical, hemato-biochemical, histological parameters and bioaccumulation in Nile tilapia (Oreochromis niloticus).

Among toxic pollutants, Mercury (Hg) is a toxic heavy metal that induces harmful impacts on aquatic ecosystems directly and human being's health indirectly. This study confirmed the in vitro magnetic potential of magnetite Nano-Particles (Fe3O4 NPs) against waterborne Hg exposure-induced toxicity in Nile tilapia (Oreochromis niloticus). We further evaluate the safety profile of Fe3O4 NPs on fish growth, hemato-biochemical, histological parameters, bioaccumulation in muscles, and economy. Magnetite nanoparticles were characterized, adsorption loading to Hg ions was investigated, and testing different concentrations of Fe3O4 NPs (0.2, 0.4, 0.6, 0.8, and 1.0 mg/L) was applied to determine the highest concentration of adsorption. An in vivo experiment includes 120 fish with an average weight of 26.2 ± 0.26 g were randomly divided into 4 equal groups, each group had three replicates (n = 30 fish/group; 10 fish/ replicate). All groups were fed on a reference basal diet and the experiment was conducted for 30 days. The first group (G1) was allocated as a control. The second group (G2) received 1.0 mg/L aqueous suspension of Fe3O4 NPs. The third group (G3) was exposed to an aqueous solution of Hg ions at a concentration of 0.025 mg/L. Meanwhile, the fourth group (G4) acquired an aqueous suspension composed of a mixture of Hg ions and Fe3O4 NPs as previously mentioned. Throughout the exposure period, the clinical signs, symptoms, and mortalities were recorded. The Hg ions-exposed group induced the following consequences; reduced appetite resulting in reduced growth and less economic efficiency; microcytic hypochromic anemia, leukocytosis, lymphopenia, and neutrophilia; sharp and clear depletion in the immune indicators including lysozymes activity, immunoglobulin M (IgM), and Myeloperoxidase activities (MPO); significant higher levels of ALT, AST, urea, creatinine, and Superoxide dismutase (SOD); histological alterations of gill, hepatic and muscular tissues with strong expression of apoptotic marker (caspase 3); and a higher accumulation of Hg ions in the muscles. Surprisingly, Fe3O4 NPs-supplemented groups exhibited strong adsorption capacity against the Hg ions and mostly removed the Hg ions accumulation in the muscles. Also, the hematological, biochemical, and histological parameters were recovered. Thus, in order to assess the antitoxic role of Fe3O4 NPs against Hg and their safety on O. niloticus, and fill the gap of the research, the current context was investigated to evaluate the promising role of Fe3O4 NPs to prevent Hg-exposure-induced toxicity and protection of fish health, which ascertains essentiality for sustainable development of nanotechnology in the aquatic environment.

Low temperature stress in a cultured fish (Piaractus mesopotamicus) fed with Pyropia columbina red seaweed-supplemented diet.

This study aimed to analyze the cold stress effects (in terms of hematology, energy reserves, and oxidative stress) in Piaractus mesopotamicus (pacú) and their mitigation by a Pyropia columbina red seaweed-supplemented diet. For this purpose, juvenile fish were fed with a control (CD) or a red seaweed-supplemented diet (RD) for 60 days, and then, the animals were exposed to a low temperature (14 °C) and a control temperature (24 °C) for 24 h. The cold shock generated an increase of hemoglobin levels in fish fed with both diets. In CD-fed fish, plasmatic triglycerides, cholesterol, and hepatic glycogen decreased after the thermal shock; meanwhile, the animals fed with RD showed decreased hepatic proteins, but increased cholesterol and hepatic glycogen. Regarding oxidative stress, antioxidant enzymes augmented their activity in the liver, intestine, and gills; meanwhile, lipid oxidative damage was observed in the liver and intestine of fish exposed to 14 °C and fed with both diets. Pacú was sensitive to cold shock, but no mitigation effects were observed in fish fed with the supplemented diet. Further research should target higher concentrations of P. columbina in supplemented diets to take advantage of this valuable resource.

Toxicity of gamma aluminium oxide nanoparticles in the Mediterranean mussel (Mytilus galloprovincialis): histopathological alterations and antioxidant responses in the gill and digestive gland.

PURPOSE: Accumulation of Gamma aluminium oxide nanoparticles γ-Al2O3 NPs significant impact on aquatic ecosystems. However, the toxicity of γ-Al2O3 NPs in aquatic organisms has been limited investigated. This study investigated histopathological changes and antioxidant responses induced by different concentrations of γ-Al2O3 NPs in Mytilus galloprovincialis. MATERIAL AND METHODS: In this study, mussels were exposed to different concentrations of 5 nm γ-Al2O3 NPs (0, 5, 20 and 40 mg/L) for 96 h under controlled laboratory conditions. Gill and digestive gland from mussels were assessed to histopathological (light microscopy, histopathological condition indices, digestive gland tubule types), SOD, CAT, GPx activities. RESULTS: Histopathological indices calculated higher, and significantly different in all exposure groups compared to the control group in gill and digestive gland (p < 0.05). Atrophic phase tubules proportion very high in 20 and 40 mg/L γ-Al2O3 NPs exposure groups. No significant changes in CAT activities in the gill and digestive gland (p > 0.05). Superoxide dismutase (SOD) activities significantly different (p ≤ 0.05) in the digestive gland from 20 mg/L γ-Al2O3 NPs exposures, and GPx activities significantly different (p < 0.05) in gill from 40 mg/L γ-Al2O3 NPs exposures. CONCLUSION: These results indicate that contamination of γ-Al2O3 NPs negatively affects the aquatic organism.

Supplementation of nano-selenium in fish diet: Impact on selenium assimilation and immune-regulated selenoproteome expression in monosex Nile tilapia (Oreochromis niloticus).

Selenium (Se), a fundamental element of nutrigenomic science in fish nutrition, was used to investigate its impact on selenoproteome expression and Se regulation in tilapia. Different concentrations (T1 - 0, T2 - 0.5, T3 - 1.0 and T4 - 2.0 mg/kg of feed) of dietary nano-Se were incorporated in the diets of monosex Nile tilapia. A total of 180 tilapia fingerlings with initial weight (15.73 ± 0.05 g) were stocked in 150 L capacity FRP tanks categorized into four diet groups with triplicate each for a feeding trial of 90 days. At the end of first, second and third months of the feeding trial, gill, liver, kidney and muscle tissues were harvested to evaluate the effect on the kinetics of Se bioaccumulation and assimilation as well as immune-regulated selenoprotein transcripts (GPx2, SelJ, SelL, SelK, SelS, SelW and Sepp1a) and their synthesis factors (SPS1 and Scly). The findings depicted that significantly (p < 0.05) higher weight gain was found in the diet supplemented with 1.0 mg/kg of nano-Se. The theory of second-order polynomial regression supported the same. The liver showed significantly (p < 0.05) higher Se accumulation and concentration factor among the harvested tissues in a different timeline. All the selected immune-regulated selenoproteins and synthesis factors in different fish tissues showed significantly (p < 0.05) up-regulation in the diet supplemented with 1.0 mg/kg of nano-Se for the second month. Therefore, the present findings suggested that the supplementation of nano-Se could be more effective for improved growth, better selenium regulation and expression of immune-regulated selenoproteins in the fish model.

Effects of dietary baicalin supplementation on growth performance, antioxidative status and protection against oxidative stress-induced liver injury in GIFT tilapia (Oreochromis niloticus).

Baicalin, a main bioactive compound of Scutellaria baicalensis, has a variety of pharmacological activities including antioxidation, anti-inflammation and hepatoprotection. However, there are few reports on these biological activities in fish. Therefore, the aim of this study was to assess the effects of baicalin on growth performance, antioxidative status and hepatoprotection in tilapia. The fish were fed on different doses of baicalin (0, 0.4, 0.8 and 1.6 g/kg diet). After feeding 60 days, parts of fishes were netted, and the blood, liver, gills and muscle tissues were collected to analyze the antioxidative effect. The remaining fishes were injected with saline or hydrogen peroxide (H2O2) for challenge test. The results showed that the specific growth rate of fish was slightly increased in three baicalin treatments, and the feed efficiency was clearly improved in 0.4 g/kg baicalin treatment. Meanwhile, the antioxidative capacity in blood, liver and/or gill was enhanced in treatments with 0.4, 0.8 and/or 1.6 g/kg baicalin. After challenge test, the pre-treatments with baicalin effectively alleviated H2O2-induced liver injury. In serum and liver, pre-treatments with 0.8 and/or 1.6 g/kg baicalin suppressed the oxidative damage induced by H2O2, as evidenced by improvement of the levels of SOD, T-AOC and GSH and the decline of MDA level. More important, pre-treatments with 0.4, 0.8 and/or 1.6 g/kg baicalin blocked the upregulation of mRNA levels of tlr1, myd88, irak4, rela, tnf-α and il-1ß in H2O2-induced liver injury. In summary, dietary baicalin supplementation could improve feed efficiency, enhance antioxidative ability and alleviate oxidative stress-induced hepatotoxicity in tilapia.

Dietary supplementation of garlic, propolis, and wakame improves recuperation in cadmium exposed Japanese medaka fish (Oryzias latipes).

Cadmium (Cd) toxic effects can overwhelm endogenous antioxidants and affect recuperation in fish. Thus, this study assessed antioxidative properties and ameliorating potentials of dietary supplemented garlic (D1), propolis (D2), and wakame (D3) on i) Cd bioconcentration, and ii) Cd induced toxicity during recuperation in Cd exposed Japanese medaka. In a 21-day exposure, control (0.0 mg Cd/L in water-C1) or Cd-treatment (0.3 mg Cd/L in water-T1) fish were fed medaka diets. Surviving fish in T1 were further depurated for 21-days and fed D1, D2, D3, or medaka diet (C2). Gill, liver, and muscle tissues were assessed weekly for Cd bioconcentration, metallothionein, (MT), superoxide dismutase (SOD), total glutathione (GSH), and lipid peroxidation (LPO). Results showed reduced antioxidant activity by significantly increasing tissue Cd and LPO, and significantly reducing SOD activity and GSH content in gill and muscle upon Cd exposure. In contrast, D1, D2, and D3 diets significantly reduced tissue Cd and LPO, while increasing contents of MT and GSH, and SOD activity, significantly. Other condition indices in D1, D2 and D3 groups were also significantly higher than those in C2 groups. In conclusion, dietary supplementation significantly increased recuperation and tissue functions in fish, in the order D1 > D2 > D3> C2.

Essential oil of Ocimum gratissimum (Linnaeus, 1753): efficacy for anesthesia and transport of Oreochromis niloticus.

This study aimed to evaluate the essential oil of Ocimum gratissimum L. (EOOG) for anesthesia and in the transport of Oreochromis niloticus. Experiment I determined the time of anesthesia induction and recovery during anesthesia of O. niloticus exposed to different concentrations of EOOG (0, 30, 90, 150, and 300 mg L-1). Based on data from Experiment I, Experiment II evaluated the effect of 0, 30, and 90 mg L-1 EOOG on blood parameters and oxidative stress immediately after anesthesia induction and 1 h after recovery. Experiment III evaluated the effect of 0, 5, and 10 mg L-1 EOOG on blood variables immediately after 4.5 h of transport of juveniles. Concentrations between 90 and 150 mg L-1 EOOG were efficient for anesthesia and recovery. The use of 90 mg L-1 of EOOG prevented an increase in plasma glucose. Other changes in blood parameters and oxidative stress are discussed. The use of 10 mg L-1 EOOG in transport increased plasma glucose and decreased hematocrit values immediately after transport. It is concluded that the use of 90 and 150 mg L-1 EOOG causes anesthesia and recovery in O. niloticus within the time intervals considered ideal. The use of 90 mg L-1 EOOG favored stable plasma glucose soon after anesthesia induction and 1 h after recovery, but caused changes in the antioxidant defense system by increasing hepatic and kidney ROS. The transport of 12 g O. niloticus for 4.5 h can be performed with concentration of 5 mg L-1 of EOOG.

Unveiling the effect of dietary essential oils supplementation in Sparus aurata gills and its efficiency against the infestation by Sparicotyle chrysophrii.

A microencapsulated feed additive composed by garlic, carvacrol and thymol essential oils (EOs) was evaluated regarding its protective effect in gills parasitized by Sparicotyle chrysophrii in Sparus aurata. A nutritional trial (65 days) followed by a cohabitation challenge with parasitized fish (39 days) were performed. Transcriptomic analysis by microarrays of gills of fish fed the EOs diet showed an up-regulation of genes related to biogenesis, vesicular transport and exocytosis, leukocyte-mediated immunity, oxidation-reduction and overall metabolism processes. The functional network obtained indicates a tissue-specific pro-inflammatory immune response arbitrated by degranulating acidophilic granulocytes, sustained by antioxidant and anti-inflammatory responses. The histochemical study of gills also showed an increase of carboxylate glycoproteins containing sialic acid in mucous and epithelial cells of fish fed the EOs diet, suggesting a mucosal defence mechanism through the modulation of mucin secretions. The outcomes of the in vivo challenge supported the transcriptomic results obtained from the nutritional trial, where a significant reduction of 78% in the abundance of S. chrysophrii total parasitation and a decrease in the prevalence of most parasitic developmental stages evaluated were observed in fish fed the EOs diet. These results suggest that the microencapsulation of garlic, carvacrol and thymol EOs could be considered an effective natural dietary strategy with antiparasitic properties against the ectoparasite S. chrysophrii.