Immune-antioxidant trait, growth, splenic cytokines expression, apoptosis, and histopathological alterations of Oreochromis niloticus exposed to sub-lethal copper toxicity and fed thyme and/or basil essential oils enriched diets.

Two experiments were conducted in this study, using 250 Oreochromis niloticus (O. niloticus) (average weight 30.28 ± 0.27 g). The first experiment was conducted to investigate the 96-h lethal concentration 50 (LC50) of copper chloride (CuCl2) using the probit analysis, seventy fish was divided into seven different concentration of CuCl2 (0, 22, 23, 24, 25, 26, and 27 mg/L), the accurate Cu concentrations were (1.23, 5.36, 6.02, 6.98, 7.05, 7.93, 8.12 mg/L Cu). The second experiment was conducted for investigating the effect of dietary supplementation with thyme (Thymus vulgaris, T. vulgaris) and sweet basil (Ocimum basilicum, O. basilicum) essential oils (TEO and BEO respectively) against sub-lethal Cu exposure (1/10 96-h LC50 of CuCl2). About 180 fish was divided into six groups in triplicate (10 fish/replicate, 30 fish/group). Group 1 (C) was kept as a control group with no Cu exposure and was fed the control basal diet. Group 2 (C-Cu) was fed the control basal diet and simultaneously exposed to 1/10 of the 96 h LC50 of CuCl2 (2.574 mg/L) as a sub-lethal concentration of Cu, where the realistic Cu concentration was 3.976 mg/L. Group 3 (TEO) and group 4 (BEO) were fed the diets fortified with 1%TEO and BEO, respectively without exposure to Cu. Group 5 (TEO-Cu) and group 6 (BEO-Cu) were fed the diets fortified with 1%TEO and 1%BEO, respectively, and simultaneously exposed to 1/10 of the 96 h LC50 of CuCl2 (2.574 mg/L). The growth and behavioral performance, immunological response and its related gene expression, antioxidant status, stress biomarker indicators, apoptosis biomarkers, and histopathological alteration were investigated. The results of the first experiment showed that the 96-h LC50 of CuCl2 in O. niloticus was 25.740 mg/L with lower and upper confidence limits of 25.152 and 26.356 mg/L, respectively. The results of the second experiment showed that sub-lethal Cu exposure induced growth retardation (lowered final body weight, total weight gain, and specific growth rate %), behavioral abnormalities (slower swimming activity and feeding performance), immunosuppression (lowered nitric oxide, complement-3, lysozyme, total proteins, albumin, and globulin), and lowering the hepatic antioxidant functions (higher MDA, and lower SOD, CAT, and GPx) in the exposed fish. Furthermore, alteration in the immune-related genes expression (down-regulation of IL-10 and TGF-ß and up-regulation of IL-1ß, IL-6, IL-8, and TRL-4), hepato-renal dysfunction (elevated ALT, AST, urea, and creatinine), and high levels of serum stress indicators (cortisol and glucose) were markedly evident. sub-lethal Cu toxicity induced significant up-regulation of apoptosis biomarkers involving, nuclear factor-κß (NF-κß), Bcl-2 Associated X-protein (BAX), meanwhile, the expression of B-cell lymphoma 2 (BCL2) and Proliferating cell nuclear antigen (PCNA) was remarkably down-regulated. In addition, apoptosis was also evident by histopathological investigation of branchial, hepatic, and renal sections. TEO and/or BEO dietary supplementation mitigate the destructive impacts of sub-lethal Cu exposure in O. niloticus, depending on the results of our study, it could be concluded that TEO and BEO with a 1% dietary level could be a promising antioxidant, immunostimulant, anti-stress factors, and anti-apoptosis mediators against heavy metal contaminants (Cu) in O. niloticus, providing a solution to the problem of aquatic bodies pollution, consequently aiding in the development of aquaculture industry.

Growth, hemato-biochemical, immune-antioxidant response, and gene expression in Nile tilapia (Oreochromis niloticus) received nano iron oxide-incorporated diets.

Nanotechnology has recently played a key role in tackling many aquacultures issues. Hence, the present study targets the evaluation of dietary inclusion of nano iron oxide (nFe2O3) on growth performance, hematology, immune-antioxidant responses, ionic regulation and expression of related genes in Nile tilapia (Oreochromis niloticus). Fish were fed supplementary nFe2O3 at rates of zero (control), 0.5, and 1 g/kg diet for 30 days. Obtained data demonstrated that nFe2O3 significantly (P < 0.05) augmented growth performance (final weight and length, body mass gain, specific growth rate, feed conversion ratio, and length gain rate). Hematological picture {RBCs, Hb, MCV, MCH and MCHC, and leukocytes interpretations (WBCs and monocytes)}; and biochemical indexes including (AST and ALT; total protein; and glucose, and cortisol) were significantly (P < 0.05) improved in nFe2O3 supplemented groups. Plasma ionic concentration was also altered with nFe2O3 supplementation, and 1g nFe2O3 revealed the most marked increase in plasma (Na+) potassium (K+) levels. Similarly, IgM, nitrous oxide (NO), and lysozyme activity, plus superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities showed a remarkable improvement in 1g nFe2O3 group compared to the control. Expression of Insulin-Like Growth Factor-1 (IGF-1) and interleukin 1-ß (IL-1ß) genes were significantly up-regulated in nFe2O3 supplemented groups. Briefly, dietary nFe2O3 inclusion had enhanced properties on growth; hemato-biochemical; immune, antioxidative profiles; and related genes expression of O. niloticus, with a recommended concentration of 1g nFe2O3.

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.

The effect of density on sex differentiation, sexual dimorphism, stress, and related gene expression in yellow perch.

A 180-day experiment was conducted to evaluate the effects of density on sex differentiation, sexual dimorphism, cortisol level, and stress related gene expression. Yellow perch, Perca flavescens, with initial mean body weight of 0.03 ± 0.001 g were reared in three different stocking densities: 1, 2, and 4 fish/L, termed as low (LD), moderate (MD), and high (HD) density, respectively, in a flow-through tank system. Results showed no significant differences in sex ratio in all density groups compared to normal population 1:1, and sexual size dimorphism (SSD) appeared when male and female were as small as the mean size reaching 11.5 cm and 12.3 cm in total length (TL) or 13.2g and 16.9g in body weight (BW), respectively. This female-biased sexual growth dimorphism was more pronounced in LD, although it was observed across all density groups. A significantly higher condition factor (K) of females than males in the LD group, and significantly higher R values of LD and MD than HD with the length/weight (L/W) linear relationships in females, were observed. Parallelly, fish reared in LD showed significantly higher mean body weight than those in the MD and HD groups, but there were no significant differences between the MD and HD. Similar results were also observed in all the other parameters of weight gain, specific growth rate (SGR), condition factor (K), and survival. These findings suggested that high density not only affected growth itself, but also affected SSD, growth trajectory or body shape, and general wellbeing in fish, especially in females. There were no significant differences in gonadosomatic index (GSI) and viscerosomatic index (VSI) among all the density groups; however, the hepatosomatic index (HSI) of LD was significantly higher than MD and HD, suggesting high density affected liver reserves or functions. Physiologically, plasma cortisol level was significantly highest in the LD among all groups, followed by MD, and lowest in HD. At the molecular level, the expression of the 70-kDa heat shock protein (Hsp70), glutathione peroxidase (GPx), and superoxide dismutase (SOD) genes involved in cellular stress were significantly upregulated in the HD group. The most significantly downregulated expression of these genes was consistently observed in the MD when compared to the LD and HD groups. In conclusion, increasing density induced chronic stress in yellow perch without affecting sex differentiation, but negatively affected expression of stress-related genes and mobilization of liver reserve, resulting in poorer wellbeing and reduced SSD, growth, and survival.

Heavy metals bioaccumulation in marine cultured fish and its probabilistic health hazard.

In aquacultures, heavy metals could be accumulated in fish tissues from natural and human-related sources depending on different factors. This study aims to estimate the level of bioaccumulation of heavy metals in cultured Gilt-head sea bream Sparusaurata. In this regard, heavy metals concentrations were measured in both water and fish musculature that were collected from a private fish farm in Kafr ElSheikh, Egypt. Regarding the water samples, heavy metals were within the permissible limits with exception of Cd, Cu, and Zn. In fish musculature, all heavy metals were within the WHO/FAO permissible limits. The bioaccumulation factor (BAF) indicated that mostly all heavy metals accumulation in the Gilt-head sea bream musculature decreased with time which may be correlated with the increase in water pH, calcium, and other cations concentrations. The hazard index (HI) calculations indicate no adverse health effects of heavy metals on humans through daily fish consumption so far. However, health risks are not negligible making the regular monitoring of metal contaminants in the studied area a necessity.

Probiotic Zado® (Ruminococcus Flavefaciens) boosts hematology, immune, serum proteins, and growth profiles in Nile tilapia (Oreochromis niloticus).

Probiotics application in aquaculture could be a key solution to enhance the overall immune and growth indicators of cultured fish. Several bacteria have demonstrated encouraging results as probiotics for fish. The current study evaluated the possible effects of Zado® (Ruminococcus Flavefaciens 28 × 104 CFU) dietary incorporation at 1 and 2 g/kg diet for 6-weeks on growth, hematological profile, immune performance, the biochemical, and anti-oxidative profiles of Oreochromis niloticus. Sampling was performed at the end of the third and sixth week. Fish fed with Zado® enriched diets showed (P < 0.05) significantly improved hematologic (MCHC, MCH, MCV, and PCV and RBCs count) and leukocytic readings (WBCs, monocytes and lymphocytes). The immune (phagocytosis, lysozyme U/ml, and nitric oxide pmol/ml) parameters were (P < 0.05) markedly increased in Zado® incorporated groups. Biochemical parameters (globulin, albumin and total proteins; AST and ALT) levels showed significant (P < 0.05) improvement at three and six weeks in Zado® groups. Serum glucose concentration was significantly higher in Zado® groups at three weeks, while was only higher for 2 g/kg Zado® at six weeks. Also, cortisol level was lower in both Zado® groups at three weeks, while was only lower for 1 g/kg Zado® at six weeks. In addition, antioxidants Gpx, SOD, and CAT were (P < 0.05) significantly higher in Zado® treatments, while pro-oxidant MDA was (P < 0.05) significantly decreased. Moreover, growth performance was also (P < 0.05) markedly boosted in Zado® incorporated groups compared to the control. Conclusively, our results demonstrated that Zado® probiotic is a safe alternative for O. niloticus with beneficial effects on hematological parameters, immune, biochemical, antioxidants, and growth profiles.

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.

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.