Electrical Characterization of Epoxy Nanocomposite under High DC Voltage.

This work studies the direct current breakdown characteristics of unfilled epoxy and epoxy nonconductive nanocomposites (SiO2,MgO and Al2O3). It also examines the variation of electrical properties in epoxy nanocomposites. The novel aspect of this study is that the samples of Epoxy nanocomposite were exposed to high voltages of up to six kilo volts for three hours using field electron microscopy under high vacuum conditions (10-5 mbar). The current emitted from these samples was measured at three different intervals of time. In addition, the influence of high voltage on the permittivity, loss factor (tan(δ)), and conductivity of the epoxy nanocomposite was studied. This evaluation was conducted before and after applying the voltage at room temperature, The frequency range extends from 10-2-10-7 Hz using the Novo Control Alpha-A analyzer. Current-voltage characterization was performed through field electron microscopy. The samples were characterized by scanning electron microscopy-energy dispersive X-ray spectroscopy and Fourier Transform Infrared Spectroscopy. The unfilled epoxy exhibited structural degradation, resulting in the formation of holes when exposed to high voltages of up to six kilo volts, leading to a reduction in electrical properties. Nevertheless, the addition of nanoparticles shows a significant increase in the operational lifetime of the epoxy nanocomposite. The degree of increase in the lifetime of epoxy composite varied depending on several factors such as the type of NPs introduced and their respective sizes. The epoxy/Al2O3 nanocomposite comparing with epoxy/MgO and epoxy/SiO2 nanocomposite showed elevated resistance to direct current breakdown strength and maintaining its dielectric.

Mucuna pruriens seeds extract boosts growth, immunity, testicular histology, and expression of immune-related genes of mono-sex Nile tilapia (Oreochromisniloticus).

Nutraceuticals have received increased attention in sustainable aquaculture. Consequently, the present study aimed to evaluate the dietary effects of Mucuna pruriens (MP) seed extract on growth performance, immune status, hepatic function, biochemical profiles, gonadal histology, and expression of immune-related genes in mono-sex Nile tilapia (Oreochromis niloticus). Fish were allocated into four groups and received MP at rates of 0 (control), 2, 4, and 6 g/kg diet, respectively, for 90 days. The results revealed that MP significantly (P<0.05) modulated growth performance (specific growth rate, final length, and length gain rate, body mass gain, and feed conversion ratio), lysozyme activity, and liver enzymes (AST, ALT). However, a non-significant effect on nitric oxide (NO) or immunoglobulin M (IgM) levels was detected, whereas the dietary inclusion of MP had a hypoglycemic effect. In terms of plasma globulin, albumin, globulin/albumin ratio, and cortisol, the MP receiving groups showed insignificant difference (P<0.05) when compared to controls, except for the 2 g MP-supplemented group. The lower inclusion concentration of MP (2 g/kg diet) demonstrated the best result (P < 0.05) for gonadosomatic index (GSI) and plasma testosterone level that was consistent with the histological findings reflecting an improvement in the testicular development compared with the control group. Expressions of complement component (C5) and interleukin 1-ß (IL-1ß) genes were significantly up-regulated in MP receiving groups. In conclusion, M. pruriens can be used as a safe natural economic feed additive and a low inclusion level of 2 g/kg diet is recommended to improve growth, enhance immunity, maintain liver functioning, improve testicular development, and to modulate immune-related genes in the mono-sex O. niloticus.

Chitosan nanoparticles alleviated endocrine disruption, oxidative damage, and genotoxicity of Bisphenol-A- intoxicated female African catfish.

Bisphenol-A (BPA) is widely used in production of plastic products. It can reach the ecosystems affecting aquatic organisms most likely fishes. The purpose of this study was to study the toxic effects of BPA on the biochemical variables and oxidative stress in female African catfish, Clarias gariepinus and to estimate the protective role of chitosan nanoparticles (CSNPs) against BPA toxicity. Five groups in triplicates of fish were divided as follows: group I was control, group II was treated with CSNPs (0.66 ml/L), group III was exposed to BPA (1.43 µg/L), group IV was treated with BPA (1.43 µg/L) plus CSNPs (0.33 ml/L), and group V was treated with BPA (1.43 µg/L) plus CSNPs (0.66 ml/L) for 30 days. Blood and liver tissue samples were collected at the end of experiment for the biochemical and oxidative stress biomarkers analyses. Results exhibited that serum Follicle Stimulating Hormone (FSH) and 17-ß Estradiol (E2) were significantly decreased in female catfish. While, serum Testosterone (T.) and Luteinizing Hormone (LH) were increased after exposure to BPA. Marked increment in superoxide dismutase (SOD) and malondialdehyde (MDA) levels of hepatic tissue of catfish exposed to BPA. Furthermore, significant reduction in hepatic catalase (CAT), glutathione peroxidase (GSH-px), total antioxidant capacity (TAC), reduced glutathione (GSH), and glutathione S-transferase (GST) levels were decreased significantly in BPA-exposed catfish compared to the control group. However, administration of female C. gariepinus with the low and high doses (0.33 ml/L and 0.66 ml/L) of CNPs restored the biochemical parameters to be close to the normal values of the control group and also, reduced oxidative stress induced by BPA toxicity. This improvement was evident in fish administrated with the high CSNPs dose (0.66 ml/L) compared to catfish exposed to BPA in group (III). Furthermore, the percentage of hepatic DNA damage was detected in group III exposed to BPA alone. However, it was declined after co- administration with both the low and high doses of CSNPs. The study has revealed that treatment with CSNPs has antagonistic functions against the toxicity of BPA in female African catfish.

Nano spirulina dietary supplementation augments growth, antioxidative and immunological reactions, digestion, and protection of Nile tilapia, Oreochromis niloticus, against Aeromonas veronii and some physical stressors.

The current study evaluated the effects of nano delivery of Spirulina platensis on growth performance, digestive enzymes, and biochemical, immunological, and antioxidative status, as well as resistance to Aeromonas veronii and some physical stressor challenges in Nile tilapia, Oreochromis niloticus. Three experimental fish groups (n = 270) with mean weights of 26 ± 0.30 g and mean lengths of 10 ± 0.5 cm were used; the first additive-free basal diet served as the control group, whereas the following two groups were supplemented with spirulina nanoparticles (SPNP) at 0 (control), 0.25, and 0.5%/kg diet for 4 weeks. Following the feeding trial, fish were challenged with hypoxia, cold stresses, and pathogenic bacteria (A. veronii) infection (9 × 108 CFU/ml). SPNP supplementation, especially 0.5%, (p < 0.05) significantly increased growth performance (specific growth rate % day-1, feed conversion ratio, and length gain rate %), immunological (plasma lysozyme and liver nitrous oxide) antioxidants (superoxide dismutase, catalase, and glutathione peroxidase in liver), biochemical (aspartate aminotransferase, alanine transaminase, glucose, and cortisol concentrations in plasma) assays, and digestive enzymes (lipase and amylase in plasma). The expression of liver's heat shock protein 70 (HSP70) and interleukin 1, beta (IL-1ß) genes showed a significant upregulation outline of 0.5% SPNP > 0.25% SPNP > 0% SPNP compared with the control. Protection in the incorporated fish groups exposed to A. veronii was 100% compared with the control group, which showed 50% cumulative mortalities. In conclusion, dietary SPNP supplementation improved growth performance, antioxidant activity, immune response, digestive enzymes, related gene expression, and resistance of Nile Tilapia to hypoxia, cold, and A. veronii infection. Thus, SPNP could be used as a natural therapy for controlling those stressors.

Astragalus membranaceus nanoparticles markedly improve immune and anti-oxidative responses; and protection against Aeromonas veronii in Nile tilapia Oreochromis niloticus.

The effects of dietary administration of Astragalus membranaceus nanoparticles (ANP) on immune and anti-oxidative responses, growth performance and disease resistance of Oreochromis niloticus were evaluated in the present study. Fish were divided into three groups and received the ANP at rates of 0 (control), 1, and 2%/kg diet for four weeks. After the four-week feeding trial, three fish from each replicate were sampled for immune and anti-oxidative responses evaluation, ten fish from each group were challenged with A. veronii, and nine fish from each group were subjected to cold and hypoxia challenges. It was obvious from the results that ANP significantly enhanced lysozyme activity and nitrous oxide (NO) activities, as well as improved superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities. Also, aspartate aminotransferase, alanine transaminase, glucose, and cortisol measurements showed significantly lower levels in incorporated groups compared to the control. Growth performance; and amylase and lipase digestive enzymes activities also showed markedly improved results. Expression of heat shock protein 70 (HSP70) and interleukin 1, beta (IL-1ß) genes were significantly upregulated throughout the entire experimental period. When challenged with A. veronii, the mortality of treated groups was significantly (P < 0.05) lower than the control. Current results proofs that dietary ANP had a synergistic effect on immune and anti-oxidative responses, growth performance and disease resistance of Oreochromis niloticus.

Expression of Hsp70, Igf1, and Three Oxidative Stress Biomarkers in Response to Handling and Salt Treatment at Different Water Temperatures in Yellow Perch, Perca flavescens.

Stress is a major factor that causes diseases and mortality in the aquaculture industry. The goal was to analyze the expression of stress-related biomarkers in response to different stressors in yellow perch, which is an important aquaculture candidate in North America and highly sensitive to handling in captivity. Three fish groups were established, each having four replicates, and subjected to water temperatures of 14, 20, and 26°C and acute handling stress was performed followed by a salt treatment for 144h at a salinity of 5 ppt. Serum and hepatic mRNA levels of heat shock protein (hsp70), insulin-like growth factor 1 (Igf1), glutathione peroxidase (Gpx), superoxide dismutase 1 (Sod1), and glutathione reductase (Gsr) were quantified at seven times interval over 144 h using ELISA and RT-qPCR. Handling stress caused a significant down-regulation in Hsp70, Gpx, Sod1, and Gsr at a water temperature of 20°C compared to 14 and 26°C. Igf1 was significantly upregulated at 20°C and down-regulated at 14 and 26°C. Salt treatment had a transient reverse effect on the targeted biomarkers in all groups at 72 h, then caused an upregulation after 144 h, compared to the control groups. The data showed a negative strong regulatory linear relationship between igf1 with hsp70 and anti-oxidative gene expressions. These findings could provide valuable new insights into the stress responses that affect fish health and could be used to monitor the stress.

A novel viral responsive protein (MjVRP) from Marsupenaeus japonicus haemocytes is involved in white spot syndrome virus infection.

A viral responsive protein (MjVRP) was characterized from Marsupenaeus japonicus haemocytes. In amino acid homology and phylogenetic tree analyses, MjVRP clustered in the same group with the viral responsive protein of Penaeus monodon (PmVRP15), showing 34% identity. MjVRP transcripts were mainly expressed in haemocytes and the lymphoid organ. Western blotting likewise showed that MjVRP was strongly expressed in haemocytes and the lymphoid organ. Immunostaining detected MjVRP within the cytosol next to the perinuclear region in some haemocytes. Experimental challenge with white spot syndrome virus (WSSV) significantly up-regulated the mRNA level of MjVRP in the M. japonicus haemocytes at 6 and 48 h. Flow cytometry and indirect immunofluorescence assays revealed that the ratio of MjVRP+ haemocytes significantly increased 24 and 48 h post-WSSV infection. These results suggest that MjVRP+ haemocytes have a supporting role in the pathogenesis of WSSV.

Saprolegniosis in Nile Tilapia: Identification, Molecular Characterization, and Phylogenetic Analysis of Two Novel Pathogenic Saprolegnia Strains.

Saprolegniosis is a fungal infection that leads to huge economic losses in tilapia aquaculture. Saprolegnia spp. are usually implicated as the etiological agents, but their identification is sometimes troublesome and confusing. In this study, two Saprolegnia strains (ManS22 and ManS33) were isolated from Nile Tilapia Oreochromis niloticus suffering from saprolegniosis. Both isolates were characterized morphologically and from internal transcribed spacer (ITS) sequence data. Additionally, both strains were tested for pathogenicity, and they were highly pathogenic and caused cumulative mortalities of 88.9% and 95.6%, respectively. Initially, the two strains were identified, by morphology of sexual and asexual stages, as members of the genus Saprolegnia. For more definitive identification and characterization, the ITS region of the ribosomal RNA genes was amplified and sequenced, and sequences were compared with other known sequences in GenBank. A phylogenetic tree constructed using the neighbor-joining method revealed that the two strains fell into two clusters within the species Saprolegnia parasitica. Cluster 1 included the ManS33 strain and cluster 2 the ManS22 strain. Cluster 1 grouped the ManS33 strain with other S. parasitica stains and shared 97-99% sequence similarity. Cluster 2 contained only the ManS22 strain and shared 93-94% similarity to several reference sequences of S. parasitica strains. Therefore, our findings suggest that ManS22 represents a newly described strain of S. parasitica. Received April 19, 2016; accepted October 27, 2016.

Feeding Glycyrrhiza glabra (liquorice) and Astragalus membranaceus (AM) alters innate immune and physiological responses in yellow perch (Perca flavescens).

The current work assessed the potential immunomodulatory and growth-promoting effects of Astragalus membranaceus (AM) and Glycyrrhiza glabra (liquorice) in Yellow perch (Perca flavescens). In this regard, fish with an average weight of 31 ± 1.0 g were divided into five groups, and fed daily with an additive-free basal diet (control); 1, 2, and 3% (w/w) Glycyrrhiza glabra, and the fifth diet was incorporated with a combination of 1% G. glabra-AM for a four-week period. Immunological, biochemical and growth parameters were measured; and sub-groups of fish were exposed to 1-week starvation. The results showed that incorporating AM and liquorice in the diet significantly improved Immunological [superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), Lipid peroxidase (LPx) and lysozyme activities], biochemical [Aspartate Aminotransferase (AST) and Alanine Transaminase (ALT) activities; and glucose and cortisol concentrations] and growth performance parameters [body mass gain (BMG), specific growth rate (SGR), length, condition factor (K) and feed conversion ratio (FCR)]. In addition, markedly up-regulated the expression of related genes [Insulin-Like Growth Factor-1 (IGF-1), Serum amyloid A (SAA), Complement Component C3 (CCC3), Alpha 2 Macroglobulin (A2M), SOD and GPx] in treated fish groups compared to the control. Conclusively, feeding AM and liquorice diets significantly increased (P < 0.05) growth performance, antioxidant and immune response profiles throughout the entire experiment, suggesting their beneficial rule as natural anti-stress agents.

Astragalus membranaceus (AM) enhances growth performance and antioxidant stress profiles in bluegill sunfish (Lepomis macrochirus).

This study was designed to assess the potential effects of Astragalus membranaceus (AM) on the growth performance and antioxidative stress response in bluegill sunfish (Lepomis macrochirus) exposed to challenging cold water temperature conditions. In this regard, fish with an average weight of 43 ± 1 g were divided into four groups and fed daily with an AM-free diet (control), and 1.5, 3, and 4.5 % (w/w) AM-incorporated diets for an 8-week period. Oxidative stress response, biochemical, and growth parameters were measured, and subgroups of fish were exposed to the outside challenging cold pond water temperature (1.6-9.9 °C) with an average of 7.0 ± 0.1 °C beyond the optimal temperature. The results showed that incorporating AM in the diet significantly improved growth performance parameters (body mass gain, specific growth rate, length, condition factor, and feed conversion ratio) and biochemicals (aspartate aminotransferase and alanine transaminase activities, and glucose and cortisol concentrations). In addition, markedly up-regulated superoxide dismutase, glutathione peroxidase, and catalase activities were observed in AM-treated fish groups over the control. Conclusively, feeding AM diets significantly increased (P < 0.05) growth performance and antioxidative stress profiles throughout the entire experiment, and this increase was much more pronounced at 8 weeks after the water temperature began to rise, which can be related to the nature of Bluegill fish as it is known to be a warm water fish. These findings are considered to be of great importance for sustainable aquaculture.

Construction and evaluation of an Edwardsiella ictaluri fhuC mutant.

Edwardsiella ictaluri is a Gram-negative facultative intracellular pathogen causing enteric septicemia in channel catfish. Iron is an essential micronutrient needed for bacterial virulence, and to acquire iron, many Gram-negative bacteria secrete ferric iron chelating siderophores. The ferric hydroxamate uptake (Fhu) system consists of four genes (fhuC, fhuD, fhuB, and fhuA), and is involved in the uptake of hydroxamate type siderophores across bacterial membranes. However, the Fhu system and its importance in E. ictaluri virulence have been uninvestigated. Here, we present construction and evaluation of an E. ictaluri ΔfhuC mutant. The E. ictaluri fhuC gene was deleted in-frame by allelic exchange, and the mutant's growth in media and virulence in catfish were determined. Our results indicated that deletion of the E. ictaluri fhuC gene did not affect the growth of E. ictaluri largely in both iron-replete and iron-depleted media. Addition of ferric iron sources into the iron-depleted medium improved the growth of both E. ictaluri ΔfhuC and wild type (WT). Catfish mortalities indicated that E. ictaluri ΔfhuC mutant was attenuated 2.05-fold compared with the parent strain. The catfish immunized with the E. ictaluri ΔfhuC mutant showed a high relative percent survival rate (97.50%) after re-challenge with the WT E. ictaluri strain. Taken together, our data indicates that the fhuC gene contributes to E. ictaluri virulence.