Pseudomonas putida infection induces immune-antioxidant, hepato-renal, ethological, and histopathological/immunohistochemical disruptions in Oreochromis niloticus: the palliative role of titanium dioxide nanogel.

BACKGROUND: Pseudomonas putida is a pathogenic bacterium that induces great losses in fishes, including Nile tilapia (Oreochromis niloticus). Currently, the application of nanomaterials in aquaculture practices has gained more success as it endows promising results in therapies compared to traditional protocols. OBJECTIVE: Therefore, the current perspective is considered the first report to assess the anti-bacterial efficacy of titanium dioxide nanogel (TDNG) against Pseudomonas putida (P. putida) in Nile tilapia. METHODS: The fish (n = 200; average body weight: 47.50±1.32 g) were allocated into four random groups (control, TDNG, P. putida, and TDNG + P. putida), where 0.9 mg/L of TDNG was applied as bath treatment for ten days. RESULTS: Outcomes revealed that P. putida infection caused ethological alterations (surfacing, abnormal movement, and aggression) and depression of immune-antioxidant variables (complement 3, lysozyme activity, total antioxidant capacity, superoxide dismutase, and reduced glutathione content). Additionally, a substantial elevation in hepatorenal biomarkers (aspartate and alanine aminotransferases and creatinine) with clear histopathological changes and immuno-histochemical alterations (very weak BCL-2 and potent caspase-3 immuno-expressions) were seen. Surprisingly, treating P. putida-infected fish with TDNG improved these variables and obvious restoration of the tissue architectures. CONCLUSION: Overall, this report encompasses the key role of TDNG as an anti-bacterial agent for controlling P. putida infection and improving the health status of Nile tilapia.

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.

Physiological Performance, Antioxidant and Immune Status, Columnaris Resistance, and Growth of Nile Tilapia That Received Alchemilla vulgaris-Supplemented Diets.

The current perspective is a pioneering trial to assess the efficacy of the dietary supplementation of Alchemilla vulgaris powder (AVP) in the diet of Nile tilapia (Oreochromis niloticus) on growth performance, blood picture, hepatic and renal biomarkers, immune status, and serum and tissue antioxidant capacity and to investigate the resistance against Flavobacterium columnare challenge. Fish (n = 360) were distributed into six groups (three replicates each) and received increasing AVP supplementation levels (0, 2, 4, 6, 8, and 10 g kg-1) for 60 days. Furthermore, fish were exposed to the bacterial challenge of a virulent F. columnare strain and maintained under observation for 12 days. During the observation period, clinical signs and the cumulative mortality percentage were recorded. The results demonstrated that the growth performance, feed conversion ratio, and hematological profile were noticeably enhanced in the AVP-supplemented groups compared to the control. The most promising results of weight gain and feed conversion ratio were recorded in the groups with 6, 8, and 10 g AVP kg-1 diets in a linear regression trend. The levels of hepatorenal function indicators were maintained in a healthy range in the different dietary AVP-supplemented groups. In a dose-dependent manner, fish fed AVP dietary supplements displayed significant augmented serum levels of innate immune indicators (lysozyme, nitric oxide, and complement 3) and antioxidant biomarkers (Catalase (CAT), superoxide dismutase (SOD), total antioxidant (TAC), and reduced glutathione (GSH) with a marked decrease in myeloperoxidase (MPO) and malondialdehyde (MDA) levels). Likewise, hepatic CAT and SOD activities were significantly improved, and the opposite trend was recorded with hepatic MDA. The highest AVP-supplemented dose (10 g/kg) recorded the highest immune-antioxidant status. Based on the study findings, we highlight the efficacy of AVP as a nutraceutical dietary supplementation for aquaculture to enhance growth, physiological performance, and immune-antioxidant status and as a natural economic antibacterial agent in O. niloticus for sustaining aquaculture. It could be concluded that the dietary supplementation of 10 g AVP/kg enhanced O. niloticus growth, physiological performance, immune-antioxidant status, and resistance against F. columnare.

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.

Dietary Salvia officinalis leaves enhances antioxidant-immune-capacity, resistance to Aeromonas sobria challenge, and growth of Cyprinus carpio.

The current perspective is a pioneer to assess the efficacy of Salvia officinalis leave powder (SOLP) on growth, intestinal enzymes, physiological and antioxidant status, immunological response, and gene expression of Common carp (Cyprinus carpio). We also looked into fish resistance after being challenged with Aeromonas sobria, a pathogenic zoonotic bacteria. Fish (N = 120) were fed four different experimental diets in triplicate for 8 weeks. The control diet (SOLP0 - without SOLP); meanwhile, the other three diets included SOLP of 2, 4, and 8 g kg-1 concentrations (SOLP2, SOLP4, and SOLP8), respectively. Findings demonstrated that fish fed SOLP4 and SOLP8 diets had better growth performance and improved digestion by noticeable enhancing lipase and amylase enzymes activity than other groups. Additionally, the antioxidant (superoxide dismutase and glutathione peroxidase) and immune activities (immunoglobulin M, nitric oxide, and antiprotease) clarified a significant increase (p < 0.05) in SOLP4 and SOLP8 groups. Enriched diets with SOLP4 and SOLP8 exhibited better expression of splenic genes (IL-1ß, IL-6, IL-10, TLR-2, and SOD), intestinal genes (Slc26a6) and (PepT1 or Slc15a1), and muscular genes (IGF-1 and SOD), while MSTN was down-regulated. After 8 weeks of the experimental trial, C. carpio challenged by A. sobria exhibited the highest cumulative mortality (66.67%), while SOLP8-dietary intervention showed the best results in enhancing the fish resistance against A. sobria by lessening mortalities to 13.33% followed by SOLP4 diet (20%). The outcomes indicate that the expression of splenic, muscular, and intestinal genes confirm the efficacy of SOLP on enhancing growth, digestion, and immune-antioxidant status, and recommend the potential use of SOLP especially at 4 g kg-1 level as a valuable natural economic diet additive in C. carpio culture for sustaining aquaculture.

Protective effects of Allium hirtifolium extract against foodborne toxicity of Zinc oxide nanoparticles in Common carp (Cyprinus carpio).

The use of nano-sized materials is increasingly growing, while consequent health and environmental risks are still disputed. On the other hand, plant extracts have been reported to improve fish general health status and enhance antioxidant capacity. Thus, the present study was aimed to assess potential effects of Allium hirtifolium extract (AHE) to fortify antioxidant responses of Common carp (Cyprinus carpio) exposed to foodborne Zinc oxide nanoparticles (ZnO-NPs). Five hundred and forty fish were randomly allocated into 18 tanks and received six diets including a basal diet (as control), basal diet incorporated with either 13 mg/kg (ZnO-25) or 26 mg/kg (ZnO-50) of ZnO-NPs, 1.5% AHE (AHE-1.5), and similar concentrations of ZnO-NPs plus AHE (ZnO-25-AHE) and (ZnO-50-AHE) for a period of 30 days. Results revealed that blood indices, stress biomarkers (glucose and cortisol), and antioxidant parameters and genes in AHE-1.5 group were significantly modulated and improved when compared to other groups (P < 0.05). In AHE-enriched groups, serum and liver tissue antioxidative parameters were enhanced as reflected in a noticeable decrease in malondialdehyde value and an increase in catalase, superoxide dismutase, glutathione peroxidase, and glutathione reductase. However, current results showed that diets incorporated with ZnO-NPs elevated the stress parameters besides a significant reduction for most measured biochemical parameters and AHE supplementation ameliorated these effects in terms of improving antioxidant parameters. In ZnO-25-AHE, and ZnO-50-AHE, the values for expression of GPx were found significantly (P < 0.05) different from that of ZnO-25 and ZnO-50. On the contrary, SOD showed a non-significant difference (P > 0.05) among control, ZnO-25, and ZnO-50-AHE, also in-between ZnO-25 and ZnO-25-AHE. The present results indicate that AHE supplementation could trigger antioxidant responses both at tissue and molecular levels suggesting its outstanding protective effects against foodborne toxicity of ZnO-NPs in Common carp.

Immune-antioxidant trait, Aeromonas veronii resistance, growth, intestinal architecture, and splenic cytokines expression of Cyprinus carpio fed Prunus armeniaca kernel-enriched diets.

Currently, the intervention of plant by-products in the fish diet has gained tremendous attention owing to the economic and high nutritious value. The current study is a pioneer attempt to incorporate the apricot, Prunus armeniaca kernel powder (PAKP) into the Common carp, Cyprinus carpio diets, and assess its efficacy on growth, digestion, intestinal morphology, immunity, antioxidant capacity, and splenic cytokines expression, besides the antibacterial role against Aeromonas veronii infection. Apparently healthy fish (N = 120) with an initial body weight of 24.76 ± 0.03g were allotted in 12 glass aquaria (60 L) and randomly distributed into four groups (triplicates, 10 fish/aquarium). The control group (PAKP0) was fed a basal diet without additives. The second, third, and fourth groups were provided PAKP diets with various concentrations (2.5 (PAKP2.5), 5 (PAKP5), and 10 g kg-1 (PAKP10)) respectively. After 60 days (feeding trial), sub-samples of the fish (12 fish/group) were intraperitoneally injected with 1 × 107 CFU mL-1 of A. veronii. Results revealed that body weight gain, feed conversion ratio, and specific growth rates were significantly augmented in the PAKP10 group in comparison to the other groups. The dietary inclusion of PAKP at all concentrations boosted the digestive capacity and maintained the intestinal morphology (average villus length, villus width, and goblet cells count) with a marked improvement in PAKP10. Moreover, fish fed on PAKP10 followed by PAKP5 then PAKP2.5 diets had noticeably elevated values of immunological biomarkers (IgM, antiprotease, and lysozyme activity) and antioxidant capabilities (the total antioxidant capacity, superoxide dismutase, and reduced glutathione) as well as significant up-regulation of immune and antioxidant-related genes (TGF-ß2, TLR-2, TNF-α, IL-10, SOD, GPx, and GSS). Fourteen days post-infection with A. veronii, the highest relative percentage survival of fish was observed in PAKP10 (83.33%), followed by PAKP5 (66.67%), and PAKP2.5 (50%). Our results indicated that a dietary intervention with PAKP could promise growth, digestion, immunity, and protect C. carpio against A. veronii infection in a dose-dependent manner. This offers a framework for future application of such seeds as a growth promotor, immune-stimulant, and antioxidant, besides an alternative cheap therapeutic antibacterial agent for sustaining the aquaculture industry.

Mooseer (Allium hirtifolium) boosts growth, general health status, and resistance of rainbow trout (Oncorhynchus mykiss) against Streptococcus iniae infection.

In large-scale aquaculture, the fast growth rate of fish is positively influenced by feed additives such as medicinal plants. This is however; infectious disease may reduce fish growth and cause devastating economic loss. The present study investigated in vitro antibacterial efficacy of Mooseer (Allium hirtifolium) extract against Streptococcus iniae and its in vivo effects on growth, biochemical parameters, innate immunity of rainbow trout (Oncorhynchus mykiss). Therefore, six experimental diets were designed to include different levels of Mooseer from zero (as control), 5, 10, 15, 20, and 25 g per kg diet respectively referred to as M1 to M5. Results from the antibacterial evaluation showed that Mooseer extract inhibits S. iniae growth with MIC and MBC values of 128 and 256 µg ml-1. Appreciable results were obtained in the groups supplemented with Mooseer. Mooseer enhanced growth performance, and modulated serum biochemical and immunological parameters (total protein, albumin, triglyceride, glucose, cortisol, cholesterol, lysozyme, Ig, ACH50, ALP, and protease activity), and liver enzymes (ALT, AST and ALP). The greatest effects were found for higher doses of Mooseer supplementation (M4 and M5). Meanwhile, results from the survival rate of fish challenged with S. iniae showed higher survival in M2 and M4 treatments. The present findings suggest the beneficial use of Mooseer in rainbow trout diet, with 20 g kg-1 inclusion as the recommended dose.

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.

Bioremediation effect of pomegranate peel on subchronic mercury immunotoxicity on African catfish (Clarias gariepinus).

The primary aim of the present study is to evaluate the highest concentrations of heavy metals (HMs) in Nile catfish (Clarias gariepinus, C. gariepinus) and water samples, as well as to investigate the efficiency of pomegranate peels (PPs) (supplemented either through water or diet) in enhancing fish immunity and counteracting the toxicity of high concentrations of HMs. Water and C. gariepinus samples were collected from two private fish farms in Al Sharkia Governorate. Mercury (Hg) showed the highest concentration (0.13 ppm). The adsorption capacity of PP was evaluated by testing different concentrations, 0.3, 0.6, 0.9, 1.2, and 1.5 g L-1, wherein 1.5 g L-1 revealed the highest Hg adsorption efficiency. The results indicated that Hg concentration was decreased with increased PP concentration until 72 h. In a trial that lasted for 60 days, 240 C. gariepinus (75.12 ± 3.12 g) were randomly divided into eight equal groups with three replicates per group. The first group (CT) served as the negative control (fish fed on a basal diet). The second group (PPW) was fed on a basal diet and supplemented with 0.3 g L-1 of PP via water. The third (PPD1) and fourth (PPD2) groups received basal diets enriched with 1 and 2 g PP powder/kg diet. respectively. The fifth group (Hg) served as the positive control that was fed on a basal diet and exposed to 0.13 ppm of Hg. The sixth (Hg + PPW), seventh (Hg + PPD1), and eighth (Hg + PPD2) groups were exposed to 0.13 ppm of Hg and received the same type of treatment as in second, third, and fourth groups. Hg exposure significantly elevated aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea, and creatinine levels, as well as oxidative stress biomarkers, including reduced glutathione (GSH) and malondialdehyde (MDA). Pomegranate supplementation through diet elevated the levels of red blood cells (RBCs), hemoglobin (Hb), packed cell volume (PCV), lysozyme, and anti-protease activity. Moreover, PP supplementation through water revealed minimum urea and creatinine levels, and the highest nitric oxide level. Moreover, Hg residue level in fish muscles noticeably decreased in the PP-treated groups. These results demonstrated the efficiency of PP supplementation (either through water or diet) in enhancing fish immunity and counteracting subchronic Hg toxicity.

Efficacy of nano zinc oxide dietary supplements on growth performance, immunomodulation and disease resistance of African catfish Clarias gariepinus.

Zinc (Zn) is an important trace element in fish diets that is required for growth, immunity and antioxidant defense mechanisms. The current study assessed the effects of both organic and nanoparticle zinc oxide (ZnO and ZnO-NPs, respectively) on growth performance, immune response and the antimicrobial effect against Pseudomonas aeruginosa in African catfish Clarias gariepinus. Fish were fed either a control diet or diets supplemented with organic ZnO at concentrations of 20 and 30 mg kg-1 or ZnO-NPs at concentrations of 20 and 30 mg kg-1. After 60 d, a subset of the fish was injected intraperitoneally with 3 × 107 CFU ml-1 of P. aeruginosa. Results showed that body weight gain, feed conversion ratio and specific growth rates were significantly increased in ZnO-NPs20 compared to all other groups. The dietary supplementation with 20 mg kg-1 of ZnO-NPs improved the antioxidant status of fish. Moreover, IgM, lysozyme and nitric oxide showed a significant increase in the fish which received the ZnO-NPs20-supplemented diet. A significant upregulation of growth and stress-related genes was seen in the ZnO-NPs20-supplemented group compared to other groups. However, there was no significant difference in the expression of immune-related genes among ZnO-NPs20, ZnO-NPs30 and ZnO30 groups. These findings highlight the potential use of nano-ZnO for improving growth performance, antioxidant status, immunological status and antibacterial activity against P. aeruginosa in African catfish.