Evaluation of different probiotics on growth, body composition, antioxidant capacity, and histoarchitecture of Mugil capito.

We investigated the dietary effects of the single application of Saccharomyces cerevisiae, Lactobacillus bulgaricus, and their combination on growth, proximate composition of whole fish body, antioxidant defense, and histoarchitecture of hapa-reared Mugil capito. Healthy fish (Fish weighed = 10.30 ± 0.10 g at first) were randomly allocated into 4 equal groups, each with three replicates. These groups were designed as follows: (1) a group fed a basal diet without probiotics (control), (2) a group fed a diet containing S. cerevisiae (4 g/kg diet), (3) a group fed a diet containing L. bulgaricus (2 g/kg diet), and (4) the last group fed a diet containing a combination of both, all for a duration of 60 days. Probiotic-treated groups showed significantly better growth and nutrition utilization than the control group. Significant differences were observed in the crude fat and crude protein contents among the groups, with the combination group exhibiting the highest levels. However, there were no significant variations in ash content across all groups. The highest hepatic antioxidant capacity (superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) enzyme activities) was observed in the combination group. Thiobarbituric acid reactive substance (TBARS) concentrations were decreased significantly in all probiotic groups, suggesting improved oxidative stress resilience in these groups. The histomorphological analysis of the hepatopancreatic tissues revealed well-arranged parenchyma, increased glycogen storage, and melanomacrophage centers in probiotic-treated groups, particularly the combined probiotics group. Furthermore, the probiotic supplementation improved the histoarchitecture of the intestinal villi compared to the control group. To put it briefly, combined dietary administration of these probiotics improved growth, body composition, antioxidant defenses, and hepatic and intestinal health in hapa-reared M. capito, highlighting their promising role in promoting welfare and productivity.

Dietary effects of Saccharomyces cerevisiae and Allium sativum on growth, antioxidant status, hepatic and intestinal histoarchitecture, expression of growth- and immune-related genes, and resistance of Oreochromis niloticus to Aeromonas sobria.

This study investigated the benefits of yeast, Saccharomyces cerevisiae and/or garlic, Allium sativum supplementation in diets of Nile tilapia with regard to growth, antioxidant status, hepatic and intestinal histoarchitecture, expression of growth- and immune-related genes, and resistance to Aeromonas sobria infection. Fish (with an initial weight of 9.43 ± 0.08 g) were allocated to twelve hapas, organized into four triplicate treatment groups defined as control (no supplementation), yeast (4 g/kg diet), garlic (30 g/kg diet), and a mixture of both. This trial continued over a 60-day feeding period. Results revealed that combined treatment (yeast + garlic) demonstrated the most promising outcomes regarding growth, with significantly higher final body weights, weight gains, and specific growth rates compared to other groups. Moreover, this combination enhanced hepatic antioxidant status, as evidenced by elevated levels of reduced glutathione and activities of catalase and superoxide dismutase enzymes, reflecting improved defense against oxidative stress. Histological assessments of the livers and intestines demonstrated structural enhancements in yeast and garlic treatments, suggesting improvements in organ health. In comparison to the control, the gene expression analyses unveiled increased expression of growth-related (igf-1 and ghr1) and immune-related (il-10, lyz, and hep) genes in the test groups, indicating a possible reinforcement of the growth and immune responses. The combined treatment also showed the highest resistance to A. sobria infection, as evidenced by improved survival rates and lower mortality compared with the other groups. These findings highlight the benefits of a combination of both yeast and garlic as a dietary supplementation regimen. In conclusion, this study suggests that the combined treatment regimen could be considered an effective strategy to promote the health and productivity of Nile tilapia under production conditions.

Dietary effects of microalga Tetraselmis suecica on growth, antioxidant-immune activity, inflammation cytokines, and resistance of Nile tilapia fingerlings to Aeromonas sobria infection.

The dietary effects of the green microalga Tetraselmis suecica (TS) on the growth, digestive enzymes, immune and antioxidant responses, genes expression, and disease resistance of Nile tilapia (Oreochromis niloticus) fingerlings were investigated. This microalga was mixed with the diet' ingredients at doses of 0.0 (the control), 5, 10, 15, and 20 g/kg diet and then fed to fish daily for 84 days. After the feeding trial, fish were experimentally challenged with Aeromonas sobria, infection and fish mortalities were recorded for another 10 days. Dietary TS significantly (p < 0.05) enhanced growth, digestive enzymes activities, and blood proteins, particularly at the level of 15 g/kg diet. Feeding the fish on 15 TS/kg feed exhibited highest mRNA expressions of GH and IGF-1 genes as well as SOD, CAT, and GPx genes compared to other TS groups. Moreover, highest levels of hepatic antioxidant and immune indices were found in the treatment of 15 g TS/kg feed. Significant downregulation of IL-1ß and IL-8 genes expression and significant upregulation of IL-10 gene expression were observed in TS-fed fish, principally in fish groups fed on 15-20 g TS/kg feed. Conversely, hepatic malondialdehyde levels, blood glucose, and the activities of transaminases (ALT and AST) were significantly (p < 0.05) decreased in fish fed with 15-20 g TS/kg diet. Serum bactericidal activity against A. sobria was significantly higher in TS-fed fish groups, and its highest levels were found in treatments of 15-20 g/kg diet. Of interest, the survival rates of fish groups fed diets with 10-20 g TS/kg feed were higher after the challenge with A. sobria infection than the control group. Accordingly, we can conclude that supplementing fish diets with a 15 g TS/kg diet enhanced the growth, antioxidant and immune activities, and resistance of Nile tilapia fingerlings to possible A. sobria infection.

Effects of L-ascorbic acid on growth, non-specific immunity, antioxidant capacity, and intestinal and hepatopancreatic histology of red swamp crayfish, Procambarus clarkii.

This study investigated the dietary effects of coated L-ascorbic acid (LA) on growth, feed utilization, survival, serum biochemical indices, immunity, antioxidant capacity, and intestinal and hepatopancreatic histology of the pre-adult red swamp crayfish. Four isoproteinous and isolipidic diets were formulated to contain several LA levels as 0, 1300, 1600, and 1900 mg/kg and designated as control (LA0), LA13, LA16, and LA19, respectively. However, the analyzed LA concentrations in diets were 0.00, 199.57, 360.45, and 487.50 mg/kg in LA0, LA13, LA16, and LA19, respectively. Triplicate treatments of crayfish (21.60 ± 0.14 g) were fed the test diets and reared in fiberglass tanks with a density of 20 individuals per each for eight weeks. Results revealed that all LA treatments had significantly enhanced growth performance compared to the control. Of interest, the LA16 treatment recorded the highest final tank biomass, biomass gain, total feed intake, condition factor, and muscle yield among the other treatments. The tank feed conversion ratio was significantly decreased in LA treatments compared to the control. Moreover, dietary LA16 and LA19 had significantly higher survival rates (93.3%) compared to (85.0%) in the LA0 group. All dietary doses of LA significantly increased serum parameters (total protein, albumin, globulin, lysozyme activity) and respiratory burst activity compared to the LA0 treatment. Dietary LA16 significantly boosted the hepatopancreatic antioxidant capacity, manifested by decreased malondialdehyde concentrations, increased catalase, superoxide dismutase, and glutathione peroxidase enzyme activities, and reduced glutathione content compared to the LA-free diet. A normal histoarchitecture of the hepatopancreatic tubules was found in all LA treatments except with some minor degenerative changes in the tubular lumen, and hepatopancreatic cells associated with enlarged nuclei were found in the LA19. However, normal intestinal histoarchitecture was found in all treatments with no recorded intestinal lesions. Of interest, the polynomial regression performed on the analyzed LA concentrations suggested that 380 mg/kg would be suitable to provide maximal biomass gain for pre-adult crayfish. In conclusion, results revealed that coated LA could enhance the growth, immunity, and antioxidant capacity of pre-adult red swamp crayfish, suggesting its potential as a functional and necessary micronutrient for crayfish diets.

Protective efficacy of dietary natural antioxidants on microplastic particles-induced histopathological lesions in African catfish (Clarias gariepinus).

Microplastic particles (MPs) are a common environmental pollutant easily ingested by fish in aquaculture. The current study evaluated the protective efficacies of some antioxidant, e.g., lycopene, citric acid, and chlorella, against the toxic effects of MP ingestion by Clarias gariepinus using histopathological biomarkers. Five experimental groups were established, a control group receiving only a standard diet, a group exposed to 500 mg/kg MP concomitant with the standard diet, and three antioxidant groups exposed to MPs plus either lycopene (500 mg/kg), citric acid (30 g/kg), or chlorella (50 g/kg) in the standard diet. After 15 days, fish were sacrificed for histological and histochemical examinations. Histological analysis of the kidney for group 2 (fed 500 mg/kg MPs alone) revealed distributed tissue dissociation, regional glomerular hypertrophy or shrinkage, melanomacrophage accumulation, and expansion of Bowman's space, while liver tissue exhibited dilation and rupture of the central vein wall, hemorrhage, cytoplasmic vacuolation, and cellular necrosis or apoptosis. Fish exposed to MPs also exhibited connective tissue fiber accumulation around renal blood vessels, renal tubules, the central hepatic vein, hepatic blood sinusoids, and serosal, muscle, and submucosal layers of the intestine. In addition, MP exposure reduced carbohydrate (mainly glycogen) contents in the brush borders and basement membranes of renal tubules, glomeruli, and intestinal tissues as well as in the cytoplasm of hepatocytes. These signs of renal, hepatic, and intestinal histopathology were fully or partially reversed by dietary lycopene, chlorella, or citric acid. Enhancing dietary antioxidants is an effective strategy for preventing MP toxicity in Clarias gariepinus in aquaculture.

Growth performance, antioxidant activities, and immunological responses of hapa-reared thinlip mullet (Liza ramada) juveniles fed on diets supplemented with spirulina (Arthrospiraplatensis).

Spirulina (Arthrospira platensis) (SP) has been utilized for a long time as a valued feed supplement because of its proteinous content and other beneficial phytochemical compounds. Herein, we investigated the influences of SP-supplemented diets on growth, body somatic indices, digestive enzymes, hepatic antioxidant activities, and immunological responses of hapa-reared thinlip mullet (Liza ramada) juveniles. Fish were assigned in six triplicate groups and were fed for consecutive 60 days on the prepared experimental diets containing varying SP levels as 0.0, 2.0, 4.0, 6.0, 8.0, and 10.0 g/kg diet and defined as control (CNT or SP0), SP2, SP4, SP6, SP8, and SP10 groups, respectively. The results indicated that dietary SP supplementation linearly and quadratically improved the fish growth performance, and the highest growth indices were found in the SP8 group. However, dietary SP supplementation did not significantly alter feed conversion ratio (FCR), survival rate (%), hepato-somatic index, and viscera-somatic index among all experimental groups. Meanwhile, digestive enzymes (lipase, α-amylase, and proteases) in the mid-intestine were also linearly and quadratically increased in all SP-fed groups, and their uppermost values were noted in the SP8 group. Hepatic antioxidants such as superoxide dismutase, catalase, and total antioxidant capacity in SP-supplemented groups were significantly elevated than the CNT group. Conversely, hepatic malondialdehyde contents were decreased significantly along with increasing dietary SP-supplementation levels. The immunological parameters such as lysozyme, respiratory burst, and alternative complement activities were significantly elevated in SP-fed groups than in the CNT group. These findings evoked that feeding SP-supplemented diets (especially at 8.0 g/kg diet) significantly promoted the growth, digestive enzymes, hepatic antioxidant status, and immunity of L. ramada juveniles.

Immunosuppressive Effects of Thallium Toxicity in Nile Tilapia Fingerlings: Elucidating the Rescue Role of Astragalus membranaceus Polysaccharides.

This study evaluated the immunotoxic effects of thallium (Tl) in Nile tilapia fingerlings and the recovery role of dietary Astragalus membranaceus polysaccharides (ASs). An 8-week experiment was designed where 180 fishes were randomly and equally assigned in triplicates into the six groups: the control group (CNT) was reared in unpolluted water and fed a commercial diet, two groups were fed a well-balanced commercial diet plus 1.5 and 3.0 g AS/kg diet (AS0.15 and AS0.30), respectively, the fourth group was exposed to a sublethal dose of Tl (41.9 µg l-1) [equal to 1/10 of 96-h lethal concentration 50 (LC50)], and the last two groups were fed 0.15 and 0.3% AS, respectively, and concurrently exposed to Tl (41.9 µg l-1) (AS0.15+Tl and AS0.30+Tl). Fish hematobiochemical parameters, serum immunity [nitric oxide, total immunoglobulin M (IgM) levels, and lysozyme activity], transcription of hepatic interferon-γ (IFN-γ), interleukin-1ß (IL-1ß), and tumor necrosis factor-α (TNF-α), and resistance to Aeromonas hydrophila (A. hydrophila) were assessed. Hematobiochemical parameters and serum immune indices were significantly decreased in the fish group exposed to sublethal Tl concentration compared to the CNT group. Furthermore, Tl exposure significantly induced overexpression of IL-1ß, TNF-α, and IFN-γ genes (4.22-, 5.45-, and 4.57-fold higher, respectively) compared to CNT values. Tl exposure also increased the cumulative mortality (%) in Nile tilapia challenged with A. hydrophila. Remarkably, the groups fed AS0.15+Tl and AS0.30+Tl significantly ameliorated all the aforementioned parameters, but did not reach CNT values. Our findings suggest the possible immunomodulating roles of dietary AS in recovering the immunotoxic effects of Tl in Nile tilapia. We can conclude that dietary AS would be useful for maintaining the immunity of Nile tilapia fingerlings.

Clinico-pathological findings and expression of inflammatory cytokines, apoptosis, and oxidative stress-related genes draw mechanistic insights in Nile tilapia reared under ammonia-N exposure and Aeromonas hydrophila challenge.

Fish diseases have a "stress-related" nature, whereas fish exposure to stressors will increase their susceptibility to infections. It was also noted that fish exposure to biotic and abiotic stressors would exaggerate the disease signs, elicit high mortalities, and cause severe economic losses. Motile aeromonad septicemia (MAS) is a major bacterial disease affecting a variety of finfish species throughout the globe and is caused by Aeromonas hydrophila. Herein, we have evaluated the impacts of ammonia-N stress and/or Nile tilapia challenge with pathogenic A. hydrophila on the clinical picture of MAS disease. Clinical signs, postmortem lesions, histoarchitectural changes, and gene transcription analysis were studied. Fish experimentally infected with A. hydrophila were exophthalmic and showed darkened skin. Moreover, opercular hyperemia, petechial hemorrhages, and gill congestion alongside dermal ulcerations were noticed in ammonia-exposed fish. On the other side, fish exposed to both stressors exhibited exophthalmia, corneal opacity, severe dropsy, and hemorrhagic dermal ulcerations. At the tissue levels, the histopathological lesions were exaggerated in the fish group exposed to ammonia stress and challenged with A. hydrophila than fish group exposed to each one alone. At the molecular levels, the mRNA expression analysis reveals significant upregulation of inflammatory cytokines such as interleukin-1 beta, CXC chemokine, and tumor necrosis factor-alpha in the kidney tissues of Nile tilapia exposed to ammonia and/or challenged with A. hydrophila. In a similar trend, the mRNA expression values of heat shock protein 70 (HSP70), oxidative stress related genes (SOD and CAT) and apoptosis-related genes (caspase 3, BAX, and cytochrome P450) were also increased in the hepatic tissues of fish exposed to singular or dual stressors. Interestingly, the highest expression levels of the above-mentioned genes were found in the fish group exposed to both stressors. Taken together, these findings indicate the occurrence of severe inflammatory and apoptotic changes in fish exposed to ammonia and infected with A. hydrophila more than each one alone. In contrast, there was a significant decrease in the expression values of the antioxidant enzyme glutathione-S-transferase (GST) in stressed fish, suggesting the occurrence of oxidative stress. This study will be helpful to draw mechanistic insights into the exposure of fish to ammonia stress and infection with A. hydrophila.

Health benefits and potential applications of fucoidan (FCD) extracted from brown seaweeds in aquaculture: An updated review.

Nowadays, the application of immunomodulators in aquaculture has become of an urgent need because of high incidence of fish and shrimp diseases. For a long time, researchers have paid great interest to find suitable, relatively economical, and environmentally safe immunostimulant products to be used either as feed or water additives to boost immunity and increase the resistance of fish and shrimp against the challenging pathogens. Probiotics, prebiotics, synbiotics, phytobiotics, herbal extracts, microalgae, macroalgae, and essential oils have been extensively evaluated. Brown seaweeds (Phaeophyceae) are a large group of multi-cellular macroalgae that are widely distributed in marine aquatic environments. They are abundant in several bioactive sulfated polysaccharides known as fucoidan (FCD). Research studies demonstrated the beneficial functions of FCD in human medicine because of its immunomodulating, antioxidant, anti-allergic, antitumor, antiviral, anti-inflammatory, and hepatoprotective effects. In aquaculture, several researchers have tested the benefits and potential applications of FCD in aquafeed. This literature review provides an updated information and key references of research studies that focused principally on using FCD in aquaculture. Its effects on growth, intestinal health, antioxidant capacity, and immune responses of several finfish and shellfish species will be discussed. This review paper will also highlight the potential efficacy and mechanisms of FCD in the modulation of toxicity signs and increasing the resistance of fish and shrimp against bacterial and viral infections. Hence, this contribution will be valuable to maintain aquaculture sustainability and to improve the health and welfare of farmed fish and shrimp.

Effects of sodium butyrate nanoparticles on the hemato-immunological indices, hepatic antioxidant capacity, and gene expression responses in Oreochromisniloticus.

Recently, nanotechnology has been greatly developed to provide the aquaculture industry with new beneficial nanomaterials to improve the health and welfare of aquatic animals. Herein, an eight-week experiment was designed to examine the dietary impacts of sodium butyrate nanoparticles (SB-NPs) on the hematological profile, blood proteins, immunological indices, antioxidant capacity, and expression analysis of cytokines and antioxidant-related genes in Oreochromis niloticus. Fish were randomly assigned into 5 experimental groups (3 replicates per group) and were fed diets supplemented with 5 levels of SB-NPs as 0.0 (control), 0.5, 1.0, 1.5, and 2.0 mg kg-1. The results revealed that supplementing diets with SB-NPs (1.0-2.0 mg kg-1) significantly elevated erythrocyte and leukocyte counts, hemoglobin concentrations, hematocrit values, total albumin, globulin, serum lysozyme activities, and total immunoglobulin M values compared with the control group. Notably, the highest levels of the parameters mentioned above were noticed in the group fed diet supplemented with 1.5 mg kg-1 SB-NPs. Moreover, dietary SB-NPs modulated the fish's antioxidant defense mechanisms, whereas there was a significant increase in hepatic superoxide dismutase, catalase, and glutathione peroxidase enzyme activities along with a significant decline in hepatic malondialdehyde concentrations in fish groups fed diets supplemented with SB-NPs (1.0-2.0 mg kg-1). A significant upregulation of antioxidant enzyme genes (gpx and sod), anti-inflammatory cytokine (il-10), and pro-inflammatory cytokines (il-1ß and il-8) were noticed in liver tissues of SB-NPs groups (0.5-1.5 mg kg-1). The highest mRNA expression folds of the above genes were recorded in the fish group fed diet supplemented with 1.5 mg kg-1 SB-NPs. In this context, we hypothesized that dietary supplementation with SB-NPs can boost the antioxidant status and immunity of O. niloticus. However, further research studies are still recommended.

Astragalus membranaceus polysaccharides modulate growth, hemato-biochemical indices, hepatic antioxidants, and expression of HSP70 and apoptosis-related genes in Oreochromis niloticus exposed to sub-lethal thallium toxicity.

A 60-day experiment was performed to assess the efficacy of dietary Astragalus membranaceus polysaccharides (ASP) in attenuation of sub-lethal thallium (Tl) toxicity in Nile tilapia. Six experimental groups (in triplicates) were designed where a fish group was raised in clean water and fed basal diet and served as control (CONT), two groups were fed the basal diet supplemented with 0.15% and 0.30% ASP (ASPL and ASPH), Tl-intoxicated group exposed to 1/10 of 96-h LC50 (= 41.9 µg/L), and two other groups were fed 0.15% and 0.30% ASP and concomitantly exposed to 41.9 µg Tl/L (ASPL-Tl and ASPH-Tl). At the end of the experiment, fish behavioral responses, clinical signs, survivability, growth, whole-body composition, intestinal digestive enzymes, serum biochemical parameters, hepatic antioxidative biomarkers, and transcription of stress and apoptosis genes were assessed. Results showed that the whole-body composition, intestinal α-amylase and protease enzymes, serum AST and blood urea levels, and hepatic GSH were not significantly different among groups (P > 0.05). The Tl-intoxicated fish group was off food, had darkened skin, showed restlessness and hyperexcitability, and high mortalities. FBW, WG, SGR and FI were significantly decreased alongside increase FCR in the Tl-exposed group. Tl exposure caused significant increases (P < 0.05) in intestinal lipase enzyme and serum indices such as ALT, creatinine, total cholesterol, triglycerides, glucose, and cortisol levels. Moreover, a significant decreases in hepatic CAT and SOD enzyme activities and significant increases in hepatic MDA contents were also noticed (P < 0.05). Furthermore, Tl exposure induced significant upregulation of hepatic HSP70 and apoptosis-related genes (p53 and caspase 3). Interestingly, dietary supplementation with ASP in ASPL-Tl and ASPH-Tl groups modulated the parameters mentioned above but still not reached the CONT values. Altogether, this study suggests that ASP could be beneficial in the modulation of sub-lethal Tl toxicity effects in Nile tilapia. Additionally, we can conclude that using natural feed supplements such as ASP in aquafeed might be necessary for maintaining the overall health performances of Nile tilapia.

Effects of GnRHa and hCG with or without dopamine receptor antagonists on the spawning efficiency of African catfish (Clarias gariepinus) reared in hatchery conditions.

Optimization of artificial reproduction is essential for minimizing genetic diversity, especially when fish are captured from their natural habitats and spawned in controlled conditions. In the present study, there was evaluation of the effects of gonadotropin-releasing hormone analogue (GnRHa) and human chorionic gonadotropin (hCG) with or without dopamine receptor antagonists such as domperidone (DOM) and metoclopramide (MET) on the spawning efficiency of African catfish (Clarias gariepinus) reared in captivity. The control group was intramuscularly (IM) injected with 1 mL of sterile saline solution. The fish specimens of the other six groups were injected IM with GnRHa or hCG, or in combination with either DOM or MET. None of the specimens had ovulations in the control group. There was the longest latency period in specimens treated with only GnRHa or hCG. There were the largest egg mass weight, fecundity, and hatchability (%) in specimens of the GnRHa + MET group. These findings indicate that GnRHa or hCG combined with dopamine receptor antagonists such as DOM and MET resulted in a marked enhancement of ovulation rate and increased the egg mass, fecundity, and hatchability of the treated C. gariepinus, and the values when there was inclusion of the MET treatment exceeded those when there was treatment with DOM.

Ginkgo biloba leaf extract improves growth, intestinal histomorphometry, immunity, antioxidant status and modulates transcription of cytokine genes in hapa-reared Oreochromis niloticus.

This study was designed to evaluate the impacts of dietary supplementation with Ginkgo biloba leaf extract (GBL) on the growth, intestinal histomorphometry, immunity, antioxidant status, and expression of cytokine genes in Nile tilapia reared in the hapas. A control diet was enriched with different GBL levels (0.0, 5.0, 7.0, and 9.0 g/kg) to form 4 experimental diets and were fed to Nile tilapia for 8 weeks. The findings illustrated that dietary GBL significantly enhanced the growth and feed utilization indices compared to those reared in the control group. A dose-dependent increase of hepatic catalase, superoxide dismutase, and glutathione peroxidase activities alongside a decline of hepatic malondialdehyde levels were recorded in GBL groups compared with the control. Serum lysozyme activity, complement C3, and immunoglobulin M levels were significantly increased in GBL groups compared with the control group. Moreover, dietary GBL maintained the normal intestinal and hepatopancreatic histological structures with a significant increase of some histomorphometric measurements of proximal, middle, and distal intestinal parts of the treated fish. Interestingly, dietary GBL supplementation significantly increased the mRNA expression of interleukin-1 beta (IL-1ß), IL-6, IL-10, tumor necrosis factor alpha (TNF-α) and interferon gamma (INF-γ) genes in the splenic tissues of treated fish over the control group. To conclude, it could be recommended to use GBL as a functional phytogenic feed additive to improve the growth, hepatic and intestinal health status, hepatic antioxidant status, and immunity of treated Nile tilapia. Besides, the second order polynomial regression revealed that 7.50 g GBL/kg diet is the optimal inclusion level to improve growth with no negative impacts on the overall health condition of treated Nile tilapia.

Copper Oxide Nanoparticles Alter Serum Biochemical Indices, Induce Histopathological Alterations, and Modulate Transcription of Cytokines, HSP70, and Oxidative Stress Genes in Oreochromis niloticus.

In the present study, fish were exposed to sub-lethal doses of CuONPs (68.92 ± 3.49 nm) (10 mg/L, 20 mg/L, and 50 mg/L) for a long exposure period (25 days). Compared to the control group (0.0 mg/L CuONPs), a significant dose-dependent elevation in blood urea and creatinine values, serum alanine transaminase, aspartate transaminase, and alkaline phosphatase enzyme activities were evident in CuONPs-exposed groups (p < 0.05). Fish exposure to 50 mg/L CuONPs significantly upregulated the transcription of pro-inflammatory cytokines (tumor necrosis factor-alpha, interleukin-1beta, interleukin 12, and interleukin 8), heat shock protein 70, apoptosis-related gene (caspase 3), and oxidative stress-related (superoxide dismutase, catalase, and glutathione peroxidase) genes in liver and gills of the exposed fish in comparison with those in the control group (p < 0.05). Moreover, varying histopathological injuries were noticed in the hepatopancreatic tissues, posterior kidneys, and gills of fish groups correlated to the tested exposure dose of CuONPs. In summary, our results provide new insights and helpful information for better understanding the mechanisms of CuONPs toxicity in Nile tilapia at hematological, molecular levels, and tissue levels.

Dietary garlic and chitosan enhanced the antioxidant capacity, immunity, and modulated the transcription of HSP70 and Cytokine genes in Zearalenone-intoxicated European seabass.

The present study was performed to evaluate the toxic effects of feed-born zearalenone (ZEN) on antioxidative status, immunity, transcriptomic responses of European seabass, and the modulating roles of dietary garlic and/or chitosan powders. Fish (30.7 ± 0.6 g) were randomly arranged in five experimental groups (in triplicates), whereas the first group was fed on the control diet only without any supplements (control), and the second group was fed on the basal diet contaminated with ZEN (0.725 mg/kg diet). Three other groups were fed on ZEN-contaminated diets and simultaneously supplemented with garlic powder (GP) (30 g/kg diet) (ZEN + GP), chitosan powder (CH) (10 g/kg diet) (ZEN + CH), and a mixture of GP and CH (ZEN + GP + CH). Fish were fed on the experimental diets thrice a day for 4 weeks. Two-way ANOVA revealed a gradual decline in serum superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities in the ZEN group reaching their lowest levels at the 4th week. Contrariwise, malondialdehyde levels were markedly higher in the ZEN group reaching their highest values at the end of the experiment. A significant decline of total immunoglobulins (P < 0.05) was observed in the serum of the ZEN group, especially after the 4th week. Moreover, significant down-regulation of interleukin-4 (IL-4) and interleukin 1 beta (IL-1ß) genes (P < 0.05) alongside significant up-regulation of tumor necrosis factor-alpha (TNF-α) and heat shock protein 70 (HSP70) genes (P < 0.05) in the liver and anterior kidney of ZEN-intoxicated group. Interestingly, dietary supplementation with GP and CH significantly attenuated ZEN-induced oxidative stress, immunosuppression, and modulated transcriptomic responses of ZEN-exposed fish. Moreover, combined dietary supplementation of both feed additives resulted in better effects than each one alone.

Effects of dietary Nannochloropsis oculata on growth performance, serum biochemical parameters, immune responses, and resistance against Aeromonas veronii challenge in Nile tilapia (Oreochromis niloticus).

The current work was planned to assess the effects of dietary microalga, Nannochloropsis oculata (NP) on the growth indices, serum biochemistry, non-specific immunity, and resistance of Nile tilapia (Oreochromis niloticus) juveniles against challenge with pathogenic Aeromonas veronii. Fish (10.21 ± 0.28 g) were randomly divided into four treatments in triplicates and were fed on diets supplemented with different levels of NP (0, 5, 10, and 15%) for eight weeks. The results showed that the dietary 5% NP significantly improved the growth parameters (final body weight, weight gain, specific growth rate, and total feed intake) (P < 0.05). Meanwhile, the feed conversion ratio was significantly decreased in NP-supplemented groups with respect to the control group. The serum protein profile (total protein, albumin, globulin, and albumin/globulin ratio) was significantly decreased by 15% NP supplemented group. Meanwhile, liver enzymes (alanine transaminase, and aspartate transaminase) activities were not significantly affected by NP dietary supplementation (P < 0.05). Serum lysozyme activity, nitric oxide (NO), and nitroblue tetrazolium (NBT) levels were significantly enhanced only in 5% NP supplemented group (P < 0.05). Interestingly, the hepatopancreatic and intestinal tissues had apparently normal histomorphology of Nile tilapia fed at 5% NP-supplemented diets. Significant upregulation of cytokines [interleukin 1beta (IL-1ß), interleukin 8 (IL-8), interferon-gamma (IFN-γ), transforming growth factor-beta (TGF- ß), and tumor necrosis factor-alpha (TNF-α)], with significant downregulation of the antioxidant gene [superoxide dismutase (SOD)] in 10% and 15% NP supplemented groups. Attractively, the relative level of protection (RLP) against challenge with pathogenic A. veronii was significantly elevated in 10% and 15% NP supplemented groups. Conclusively, the obtained results reflect the beneficial roles of dietary NP to improve growth and boost the immune responses of Nile tilapia.

Dietary Origanum vulgare essential oil attenuates cypermethrin-induced biochemical changes, oxidative stress, histopathological alterations, apoptosis, and reduces DNA damage in Common carp (Cyprinus carpio).

The study was designed to evaluate the possible protective roles of dietary Origanum vulgare essential oil (OVEO) against cypermethrin (CP)-induced serum biochemical changes and oxidative stress of common carp (Cyprinus carpio). Moreover, histopathological alterations, apoptosis, cell proliferation, and DNA damage in the gills and hepatic tissues were also assessed. Briefly, fish were allotted into six groups with three triplicates whereas a group fed on basal diet and did not exposed to CP and served as control (CTR), two groups were fed on diets supplemented with two levels of OVEO (0.5 % and 1.0 %), a group exposed to sub-lethal concentration of CP (1/10 of 96 h-LC50 = 0.4134 µg/L), and two other groups exposed to the same concentration of CP and fed on diets supplemented with both levels of OVEO (CP + 0.5 % OVEO, and CP + 1.0 % OVEO), respectively, for 30 days. CP induced significant elevation of serum alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), urea, and creatinine levels indicating hepato-renal toxicity (P < 0.05). Besides, there was a significant decrease in serum catalase (CAT) and superoxide dismutase (SOD) activities (P < 0.05). Moreover, CP induced significant histopathologic alterations in gills, anterior kidneys, and hepatic tissues with activation of apoptosis (Caspase-3) and proliferating cell nuclear antigen (PCNA). Comet assay demonstrated significant DNA damage in gills and liver tissues of the CP-exposed group. Interestingly, a significant attenuation of serum ALT, AST, ALP, urea, creatinine, CAT, and SOD levels (P < 0.05) was noticed in CP-exposed fish and concurrently fed diets supplemented with either 0.5 % or 1.0 % OVEO. Moreover, histopathologic alterations and apoptosis were significantly reduced along with a concomitant significant decrease in DNA damage (P < 0.05) which indicated the mitigation of DNA damage. Conclusively, the study showed that OVEO is an effective counteractive treatment against CP-induced damage in exposed common carp and is recommended during the formulation of fish rations.

The growth performance, antioxidant capacity, immunological responses, and the resistance against Aeromonas hydrophila in Nile tilapia (Oreochromis niloticus) fed Pistacia vera hulls derived polysaccharide.

The present study aimed at evaluating the supplementation of dietary Pistacia vera hulls derived polysaccharide (PHDP) at varying levels (0, 2.5, 5, and 10 g/kg diet) on the growth rate, digestive enzyme activity, immune response, and antioxidative capacity of Nile tilapia. After the feeding trial (60 days), fish were injected intraperitoneally with Aeromonas hydrophila, and the cumulative mortality was calculated for 10 days. The final body weight, weight gain, specific growth rate, survival rate, amylase activity, red blood cells, hemoglobin, serum total protein, and serum lysozyme activity (LZM) were significantly higher in fish fed PHDP at 5 and 10 g/kg than fish fed 0 and 2.5 g/kg diet (P ≤ 0.05). However, the feed conversion ratio and glucose levels were significantly decreased by 5 and 10 g PHDP/kg diet. Further, the protease digestive activity, serum alkaline phosphatase, and serum alternative complement (ACH50) had higher values in fish fed 5 g PHDP/kg diet than tilapia fed 0 g PHDP/kg diet (P ≤ 0.05). The serum protease and immunoglobulin (Ig) presented the highest values in fish fed 10 g PHDP/kg diet followed by fish fed 5 g PHDP/kg diet (P ≤ 0.05). The LZM, ACH50, and Ig in the mucus samples were significantly higher in fish fed 5 or 10 g PHDP/kg diet than fish fed 0 g PHDP/kg diet (P ≤ 0.05). The levels of superoxide dismutase and catalase in the liver tissue was significantly higher in fish fed 5 or 10 g PHDP/kg diet than fish fed 2.5 g PHDP/kg diet (P ≤ 0.05). On the other hand, malondialdehyde concentration was significantly lower in tilapia fed 5 and 10 g PHDP/kg diet than tilapia fed 0 and 2.5 g PHDP/kg diet (P ≤ 0.05). The lowest mortality rate and the highest relative percentage survival were in fish fed 5 g/kg followed by 10 g level after 10 days of A. hydrophila injection. Accordingly, dietary PHDP at 5-10 g/kg is recommended to improve the growth performance, antioxidative capacity, immune response, and resistance against A. hydrophila in Nile tilapia.

Dietary origanum essential oil improved antioxidative status, immune-related genes, and resistance of common carp (Cyprinus carpio L.) to Aeromonas hydrophila infection.

The use of herbal essential oils in aquafeeds is an important approach to maintain the fish health status. The origanum essential oil (OEO), extracted from Origanum vulgare, was included at levels of 0, 5, 10, 15, and 20 g per kg diet to evaluate its beneficial effects on antioxidant, immunity status, and expression of immune-related genes of common carp, Cyprinus carpio L. After 8 weeks, dietary inclusion of OEO significantly increased the activities of hepatic superoxide dismutase and catalase (P < 0.05); meanwhile, levels of hepatic malonaldehyde significantly decreased (P < 0.05). The levels of serum lysozyme activity, phagocytic activity, and phagocytic index have been linearly and quadratically increased by dietary OEO in a dose-dependent regime showing their highest levels in 15-20 g OEO/kg diet. Moreover, dietary OEO significantly upregulated the relative expression of hepatic interleukin-1beta (IL-1ß) and interleukin-10 (IL-10) genes (P < 0.05). Dietary OEO maintained the normal histomorphological criteria and density of melanomacrophage centers of spleens, and hematopoietic tissues of the anterior kidneys in all fish groups with respect to the control. After challenge with pathogenic bacterium Aeromonas hydrophila, the 10-day cumulative mortality significantly decreased (P < 0.05); meanwhile, the relative percent of survival of common carp significantly increased by OEO administration (P < 0.05) in a dose-dependent manner. The obtained results reflect the beneficial roles of dietary OEO to boost the antioxidative status and the immune responses in common carp with an optimum level of 15 g/kg diet.

Spirulina platensis mediated the biochemical indices and antioxidative function of Nile tilapia (Oreochromis niloticus) intoxicated with aflatoxin B1.

Aflatoxicosis is one of the threats that cause severe mortalities in fish farms. The dietary functional additives are a friendly approach attributed to beneficial effects on aquatic animals. The study aimed at evaluating the impact of Spirulina platensis (SP) on the biochemical indices and antioxidative function of Nile tilapia (Oreochromis niloticus) intoxicated with aflatoxin B1 (AFB1). A control diet and 3 test diets were enriched with 0% SP/0 mg AFB1/kg (control), 1% SP (SP), 2.5 mg AFB1/kg diet (AFB1), and 1% SP+2.5 mg AFB1/kg diet (SP/AFB1). The diets were supplied to three aquaria for each group twice daily at the rate of 2.5% for 30 days. The blood alanine transaminase (ALT), alkaline phosphatase (ALP), and aspartate transaminase (AST) were significantly increased by AFB1 toxicity with regards to fish fed the control and SP diets (P < 0.05). The inclusion of SP in the diet of tilapia intoxicated with AFB1 lowered the levels of ALT, AST, and ALP in comparison to fish contaminated with AFB1 without SP (P < 0.05). The total blood protein and albumin were decreased in fish contaminated with AFB1 (P < 0.05); however, the dietary SP resulted in improving the blood protein and albumin with similar levels with the control and SP diets. The urea and creatinine were increased in tilapia fed AFB1 diet without SP (P < 0.05); however, the inclusion of SP reduced the levels of urea and creatinine with similar levels with the control and SP diets. The antioxidative capacity of Nile tilapia fed SP and contaminated with AFB1 is expressed by superoxide dismutase (SOD), glutathione (GSH), and malondialdehyde (MDA) concentration. The activities of SOD and GSH were decreased by AFB1 (P < 0.05); however, dietary SP increased the SOD and GSH in fish fed AFB1. On the other hand, the concentration of MDA was increased in tilapia fed AFB1 (P < 0.05); however, SP decreased the level of MDA in fish fed AFB1. In conclusion, the application of SP in the aquafeed seems to be an innovative approach to relieve the toxic influences of AFB1 on aquatic animals.