Tea polyphenols alleviate TBBPA-induced inflammation, ferroptosis and apoptosis via TLR4/NF-κB pathway in carp gills.

The extensive application of Tetrabromobisphenol A (TBBPA) leads to the pollution of part of the water environment and brings great safety risks to aquatic animals. As a natural extract, tea polyphenols (TPs) have antioxidant and anti-inflammatory effects. Gills are one of the immune organs of fish and constitute the first line of defense of the immune system. However, it was unclear whether TPs could mitigate TBBPA-induced gills injury. Therefore, an animal model was established to investigate the effect of TPs on TBBPA-induced gills. The results indicated that TBBPA changed the coefficient and tissue morphology of carp gills. In addition, TBBPA induced oxidative stress and inflammation, leading to ferroptosis and apoptosis in carp gills. Dietary addition of TPs significantly improved the antioxidant capacity of carp, effectively inhibited the overexpression of TLR4/NF-κB and its mediated inflammatory response. Moreover, TPs restored iron metabolism, reduced the expression of pro-apoptotic factors thereby alleviating ferroptosis and apoptosis in carp gills. This study enriched the protective effect of TPs and provided a new way to improve the innate immunity of carp.

Tea polyphenols protect against Flavobacterium columnare-induced gill injury via suppression of oxidative stress, inflammation, and apoptosis in grass carp.

Flavobacterium columnare (F. columnare) is one of the deadliest fish pathogens causing bacterial gill rot disease in various freshwater fish species globally. Tea polyphenols (TPs) are an inexpensive product extracted from tea that have received much attention as a feed additive in aquaculture. The current study was designed to investigate the underlying mechanisms and protective effects of dietary TPs against F. columnare-induced gill injury via suppression of oxidative stress, apoptosis, and inflammation in grass carp. TPs were not supplemented to the diet (control) and were supplemented at 40, 80, 120, 160 or 200 mg/kg diet. The feeding experiment was carried out for 60 days, followed by a 3-Day F. columnare challenge test. The results showed that 120 mg/kg TPs in the diet exerted the following five protective effects in fish gill: (1) control gill-rot disease and improved histopathology, (2) inhibit excessive apoptosis, (3) enhance the activity of antioxidant enzymes and upregulate related gene expression via the Nrf2/Keap1 pathway, (4) increase the activity of immune enzymes, And (5) mediate inflammatory cytokine gene expression via the JAK/STAT3 pathway. Taken together, dietary supplementation with TPs is a compelling approach to protect the gill function of fish against F. columnare.

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.

Assessment of the water quality and toxicity effects on zebrafish (Danio rerio) of a stream near a phosphorus chemical plant in Guizhou Province, southwestern China.

To reveal the influence of the phosphorus chemical industry (PCI) on regional water environmental quality and safety, the water quality and ecotoxicological effects of a stream near a phosphorus chemical plant (PCP) in Guizhou Province, southwestern China, were investigated based on water samples collected from the stream. The results showed that the average concentrations of NH3-N, TN, P, F-, Hg, Mn, and Ni were 3.14 mg/L, 30.09 mg/L, 3.34 mg/L, 1.18 mg/L, 1.06 µg/L, 45.82 µg/L, and 11.30 µg/L, respectively. The overall water quality of the stream was in the heavily polluted category, and NH3-N, TN, P, F-, and Hg were the main pollution factors. The degree of pollution was in the order of rainy period > transitional period > dry period, and the most polluted sample site was 1100 m from the PCP. After 28 days of exposure to stream water, there was no significant change in the growth parameters of zebrafish. The gills of zebrafish showed a small amount of epithelial cell detachment and a small amount of inflammatory cell infiltration, and the liver tissue displayed a large amount of hepatocyte degeneration with loose and lightly stained cytoplasm. Compared with the control group, the %DNA in tail, tail length, tail moment, and olive tail moment were significantly increased (p < 0.05), indicating that the water sample caused DNA damage in the peripheral blood erythrocytes of zebrafish. The stream water in the PCI area was found to be polluted and exhibited significant toxicity to zebrafish, which could pose a threat to regional ecological security.

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.

Toxicological effect of endocrine disrupting insecticide (deltamethrin) on enzymatical, haematological and histopathological changes in the freshwater iridescent shark, Pangasius hypothalamus.

This study investigated the deltamethrin (DMN) induced harmful effects on Pangasius hypophthalmus using enzymatic activity, haematological, and histopathological changes. LC50 value was 0.021 mg/L at 96 h, and sublethal toxicity was tested for 45 days at two `concentrations (i.e., 1/5th and 1/10th of LC50). Haematological parameters and enzymatic activities significantly changed between DMN-exposed and control groups (p < 0.05). Histopathologically, both DMN doses induced liver hyperemia, hepatic cell rupture, necrosis, hypertrepheoid bile duct, shifting nuclei, vascular haemorrhage, and hepatocyte degeneration, while in gill, secondary lamellae destruction, a fusion of adjacent gill lamellae, hypertrophy, hyperplasia, adhesion, and fusion were noticed. Kidney developed melanomacrophages, increased periglomerular and peritubular space, vacuolation, decreased glomerulus, hyaline droplets in tubular cells, loss of tubular epithelium, distal convoluted segment hypertrophy, and granular layer in brain pyramid and Purkinje cell nucleus. But, limiting pesticide impacts on freshwater fish and their habitat requires a holistic, cradle-to-grave approach and toxicological studies.

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.

Revealing Natural Intracellular Peptides in Gills of Seahorse Hippocampus reidi.

The seahorse is a marine teleost fish member of the Syngnathidae family that displays a complex variety of morphological and reproductive behavior innovations and has been recognized for its medicinal importance. In the Brazilian ichthyofauna, the seahorse Hippocampus reidi is among the three fish species most used by the population in traditional medicine. In this study, a protocol was performed based on fast heat inactivation of proteases plus liquid chromatography coupled to mass spectrometry to identify native peptides in gills of seahorse H. reidi. The MS/MS spectra obtained from gills allowed the identification of 1080 peptides, of which 1013 peptides were present in all samples and 67 peptide sequences were identified in an additional LC-MS/MS run from an alkylated and reduced pool of samples. The majority of peptides were fragments of the internal region of the amino acid sequence of the precursor proteins (67%), and N- and C-terminal represented 18% and 15%, respectively. Many peptide sequences presented ribosomal proteins, histones and hemoglobin as precursor proteins. In addition, peptide fragments from moronecidin-like protein, described with antimicrobial activity, were found in all gill samples of H. reidi. The identified sequences may reveal new bioactive peptides.

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.

Evidence for the Presence of Hyphae and Fruiting Body Calcium Oxalate Crystallites in the Split Gill Medicinal Mushroom Schizophyllum commune (Agaricomycetes).

Biomineralization is the phenomenon by which organisms form crystals. Studies have shown that many fungi can biomineralize, producing calcium oxalate crystals on their hyphae and fruiting body. Schizophyllum commune is a medicinal and edible fungus found worldwide, however, studies on biomineralization in this fungus are limited. Here, samples of Sch. commune fruiting bodies were collected from three different provinces in China and hyphal cells were cultured. Using light microscopy, FE-SEM, and EDAX, we identified crystals on the fruiting body and mycelium of each strain and analyzed their morphological characteristics and ion content. These data demonstrate that biomineralization occurs in Sch. commune in nature as well as during subsequent in vitro culture.

Influence of TiO2 nanocomposite UV filter surface chemistry and their interactions with organic UV filters on uptake and toxicity toward cultured fish gill cells.

Aquatic environments have been found to be contaminated with a variety of inorganic and organic UV filters. This includes novel nano-sized titanium dioxide (TiO2) composite particles, which have been increasingly developed and incorporated into commercial sunscreens in recent years. So far, relatively little is known about the effects of this novel class of UV filters on aquatic life. Therefore, this study aimed to determine and compare the toxicity of three such nanoparticulate TiO2 UV filters with different surface coatings, namely Eusolex® T-Avo (SiO2-coated), T-Lite™ SF (Al(OH)3/PDMS-coated), and Eusolex® T-S (Al2O3/stearic acid-coated) either alone, or in the presence of selected organic UV filters (octinoxate, avobenzone, octocrylene), toward fish using RTgill-W1 cell cultures as an in vitro experimental model. Besides standard exposure protocols, alternative approaches (i.e., exposure to water accommodated fractions (WAFs), hanging-drop exposure) were explored to account for nanoparticle (NP)-specific fate in the medium and obtain additional/complementary information on their toxicity in different conditions. The AlamarBlue, CFDA-AM and Neutral Red Retention (NR) assays were used to measure effects on different cellular endpoints. Transmission electron microscopy (TEM) was used to examine NP uptake. Our results showed that none of the TiO2 NP UV filters were cytotoxic at the concentrations tested (0.1-10 µg/mL; 24 h) but there were differences in their uptake by the cells. Thus, only the hydrophilic T-AVO was detected inside cells, but the hydrophobic T-Lite SF and T-S were not. In addition, our results show that the presence of NPs (or the used dispersant) tended to decrease organic UV filter toxicity. The level of combination effect depended on both NP-type (surface chemistry) and concentration, suggesting that the reduced toxicity resulted from reduced availability of the organic UV filters due to their adsorption to the NP surface. Thus, mixtures of TiO2 NP UV filters and organic UV filters may have a different toxicological profile compared to the single substances, but probably do not pose an increased hazard.

Seeds of Cucurbita maxima and Carica papaya are effective for controlling monogeneans in the gills of Leporinus macrocephalus.

This study was carried out to evaluate the anthelminthic efficacy of seeds of Cucurbita maxima and Carica papaya for controlling monogeneans in the gills of Leporinus macrocephalus, besides hepatosomatic and splenosomatic index and condition factor of host. The fish were fed with seeds of C. maxima or C. papaya for seven days, and these treatments did not cause any mortality among them. Jainus leporini, Urocleidoides paradoxus, Urocleidoides eremitus and Tereancistrum parvus were the monogeneans found, and their prevalence in fish fed with seeds of C. papaya was 100%, while in fish fed with C. maxima the prevalence was 42.8%. Fish fed with seeds of C. papaya showed decreased in intensity and abundance of monogeneans, while fish fed with seeds of C. maxima presented decreased in abundance. Feeding of L. macrocephalus with seeds of C. maxima or C. papaya had efficacy of 69.6 and 67.8%, respectively. The hepatosomatic index of fish fed with seeds of C. maxima or C. papaya was not affected by the treatments. However, the splenosomatic index and condition factor of fish fed with C. maxima seeds decreased. Seeds of C. maxima and C. papaya may be used for controlling monogeneans of L. macrocephalus in fish farming.

Effect of potassium supplementation on osmoregulatory and stress response of Pangasianodon hypophthalmus (Sauvage, 1878) with Spinacia oleracea L. in aquaponics.

This study evaluated the effect of different potassium supplementation dosages on the physiological responses of Pangasianodon hypophthalmus reared in an aquaponic system with Spinacia oleracea L. for 60 days. The system comprised of a rectangular fish tank of 168 l capacity (water volume = 100 l) with nutrient film technique (NFT)-based hydroponic component with fish to plant ratio of 2.8 kg m-3 : 28 plants m-2 in all the treatments. The osmoregulatory and stress parameters of P. hypophthalmus at four different potassium dosages of T1 (90 mg l-1 ), T2 (120 mg l-1 ), T3 (150 mg l-1 ) and T4 (180 mg l-1 ) were compared with C (control, 0 mg l-1 ) to examine the potassium level to be applied to aquaponics. The water quality parameters and fish production were found to have no adverse impact due to potassium supplementation. The spinach yield during two harvests, i.e., before and after potassium supplementation, revealed that the yield was significantly higher (P < 0.05) after supplementation with the highest yield in T3 and T4. The osmoregulatory parameters such as plasma osmolality, Na+ , K+ ATPase activity in gill and plasma ionic profile (Cl- , Ca2+ and Na+ ) showed an insignificant variation (P > 0.05) between control and treatments except for higher plasma potassium concentration (1.98 ± 0.19 mmol l-1 ) in T4. The stress and antioxidant enzyme analysis exhibited significantly higher plasma glucose and superoxide dismutase (SOD) activity in gill and liver in T4, whereas cortisol and catalase showed an insignificant difference (P > 0.05). The experimental findings demonstrated that the potassium dosage up to 150 mg l-1 could be suggested as optimum for P. hypophthalmus and spinach aquaponics without impairing the health and oxidative status of P. hypophthalmus.

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.

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.

New Insight on the Immune Modulation and Physical Barrier Protection Caused by Vitamin A in Fish Gills Infected With Flavobacterium columnare.

In this study, we have investigated the influence of vitamin A on gill barrier function of grass carp (Ctenopharyngodon idella) infected with Flavobacterium columnare. The fish were fed different concentrations of vitamin A diets for 10 weeks and then infected with F. columnare by immersion. We observed that optimal vitamin A significantly prevented gill rot morbidity in fish infected with F. columnare. Further investigations revealed that vitamin A boosted the gill immunity by increasing the contents of complements (C3 and C4), activities of acid phosphatase (ACP) and lysozyme, mRNAs of ß-defensin-1, liver-expressed antimicrobial peptide 2A and 2B (LEAP-2A and LEAP-2B), hepcidin, and anti-inflammatory cytokines like transforming growth factor ß1 (TGF-ß1), TGF-ß2, interleukin-10 (IL-10), and IL-11. It also enhanced the levels of various related signaling molecules including inhibitor protein κBα (IκBα), target of rapamycin (TOR), and ribosome protein S6 kinase 1 (S6K1) but downregulated the expression of pro-inflammatory cytokines including IL-1ß, IL-8, tumor necrosis factor α (TNF-α), and interferon γ2 (IFN-γ2) and related signaling molecules including nuclear factor κB p65 (NF-κB p65) (rather than NF-κB p52), IκB kinase ß (IKKß), IKKγ (rather than IKKα), eIF4E-binding protein 1 (4E-BP1), and 4E-BP2 mRNA levels in fish gills. In addition, dietary vitamin A markedly lowered the concentrations of reactive oxygen species (ROS), malondialdehyde (MDA), and protein carbonyl (PC), increased both the activities and mRNAs of copper/zinc superoxide dismutase (Cu/ZnSOD), MnSOD, glutathione transferases (GSTs), glutathione peroxidase (GPx), and glutathione reductase (GR) associated with upregulation of NF-E2-related factor 2 (Nrf2) mRNAs and downregulation of Kelch-like-ECH-associated protein (Keap1a) and Keap1b mRNAs. Moreover, vitamin A decreased the mRNAs of different apoptotic mediators [caspases 8, 9, 3 (rather than 7)] associated with downregulation of signaling molecule p38 mitogen-activated protein kinase (p38MAPK) mRNAs in fish gills. Besides, vitamin A promoted tight junction (TJ) complex mRNAs [including claudin-b, -c, -3, -7, -12, occludin, and zonula occludens-1 (ZO-1)] that have been linked to the downregulation of myosin light chain kinase (MLCK) signaling. Taken together, the current study demonstrated for the first time that vitamin A markedly enhanced gill health associated with immune modulation and physical barrier protection. Based on protecting fish against gill rot morbidity, ACP activity, and against lipid peroxidation, optimum vitamin A concentrations in on-growing grass carp (262-997 g) were found to be 1,991, 2,188, and 2,934 IU/kg diet, respectively.

The Effect of Zinc Concentration on Physiological, Immunological, and Histological Changes in Crayfish (Procambarus clarkii) as Bio-indicator for Environment Quality Criteria.

The release of heavy metals to the environment increased dramatically with industrialization and rapid economic development, and they have accumulated in aquatic organisms. The current study aimed toe valuate the physiological, immunological, and histological changes of crayfish (Procambarus clarkii) as bio-indicator for water quality. Crayfishes of the filed study group were collected from a polluted area (Rosetta branch, Egypt), where the highest concentration for heavy metals in water was for zinc (Zn). Besides the field study group, other crayfishes were exposed to different doses of ZnSO4 (0, 203, and 406 mg L-g) corresponding to Zn concentration (0, 46.03, and 92.06 mg L-1) respectively in aquariums for consecutive 4 days. Heavy metal concentrations in field water sample were arranged as follows: Zn > Fe> Pb > Cu and Mn > Ni > Co > Cd. The result revealed that Zn bioaccumulation increases significantly with the increase of water Zn concentration among the tested groups compared to the control group, where the highest bioaccumulation in all studied tissues (hepatopancreas, gills, and muscles) was observed in the field group and Zn high-dose group. Also, there was a significant increase in the levels of hemolymph uric acid, urea, creatinine, glucose, aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase. Their highest concentrations were observed in the Zn high-dose group and the field group, while the levels of total protein, albumin, and cholesterol showed a significant decrease among the tested groups as compared with the control group. Their determined lowest concentrations were in the Zn high-dose group and field group. Among tested groups, total hemocytes and granulated hemocytes decreased significantly while hyaline hemocytes increased as compared with the control group. Histological damages were observed in hepatopancreas, gills, and muscles in the field and Zn groups. The present study showed that exposure to Zn caused physiological and histological changes in Procambarus clarkia. We assumed that Procambarus clarkia could be used as a sensitive bioindicator for monitoring water quality criteria.