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Nowadays, nanomaterials enter high numbers of daily used products and drug manufacture. A nanocomposite of vitamins C (VC) and vitamin E (VE) with chitosan as a vehicle and protector was used in a comparative eight-week feeding study, Nile tilapia weighing 31.2 ± 0.36 g distributed in seven groups and fed (G1) basal diet, (G2) bulk VC, (G3) VC- nanoparticles (NPs), (G4) bulk VE, (G5) VE-NPs, bulk VCE (G6), and (G7) VC plus VE (VCE)-NPs, respectively. The Nile tilapia-fed nanocomposite vitamins had significantly higher growth performance compared to the control; VCE-NPs had the superiority among tested supplementations where total weight gain (63.6 g), daily weight gain (1.13 g), relative growth rate (206.1%) with lower feed conversion rate (1.6) and insignificant feed intake (101.5 g). Overall, the level of liver enzymes was significantly decreased in fish serum after eight-week nanocomposite supplementation, and dietary VCE-NPs caused a significant reduction of serum AST (18.45 IU/L) and ALT (14.77 IU/L) compared to the control 25.5 IU/L and 17.6 IU/L, respectively. Fish fed dietary VCE-NPs, VC-NPs, and VE-NPs had significant enhancement of RBCs 4.2 × 10 6 /μL, 3.8 × 10 6 /μL, and 3.55 × 10 6 /μL; WBCs 46.15 × 10 3 , 42.9 × 10 3 , and 44 × 10 3 /μL, respectively, Also TP was significantly higher 6.38 g/dL in VCE-NPs group compared to the control and the other treatments. Over all, the dietary nanocomposite vitamins boost the innate immunity of the experimental Nile tilapia, the oxidative burst activity (OBA), phagocytic activity (PA), phagocytic index (PI), and serum antibacterial (SAA) were significantly increased compared to those received bulk vitamins and the control. The activity of antioxidant biomarkers in fish serum including glutathione peroxidase (GPx), catalase (CAT), superoxide dismutase (SOD), total antioxidant capacity (TAC), glutathione reductase (GR), and myeloperoxidase (MPO) showed a rise in the serum of Nile tilapia received nano- and bulk-form of VC and VCE compared to the control and both forms of VE. Furthermore, the level of malondialdehyde (MDA), reduced glutathione (GSH), and oxidized glutathione (GSSG) were significantly increased in the fish serum following the trend of antioxidants enzymes. In conclusion, a dietary nanocomposite of vitamin C and vitamin E enhanced Nile tilapia's growth performance and feed utilization. It could also improve health status and immune response. The values of antioxidant biomarkers indicated that the nanocomposite could help the fish body scavenge the generated reactive oxidative species (ROS).

This study evaluated and compared the effectiveness of Bacillus species probiotics and formalin-killed bacterin to control vibriosis caused by Vibrio anguillarium in European eel ( Anguilla anguilla ) elvers. Fish were allocated into four triplicate groups defined as follows: a control non-vaccinated (CON) group that fed on a basal diet without additives, two groups fed on diets supplied with Bacillus species probiotics (PRO; Sanolife PRO-F) in a dose of 0.5 g /kg diet (PRO-F1) and 1.0 g /kg diet (PRO-F2), and the fourth group was vaccinated with immersion formalin-killed bacterin and got a booster vaccination at 14th day (VACC). The experiment lasted for 28 days. In the end, the immune responses, including antibody titers, lysozyme activities (in serum and skin mucous), and bactericidal activity, were evaluated. The relative percent survival (RPS) after intraperitoneal challenge with V. anguillarium was calculated after an additional 10-day observation period. A one-way ANOVA followed by Dunnett’s multiple comparison test was used to compare the experimental groups (VACC, PRO-F1, and PRO-F2) with the CON group. The results revealed significantly higher antibody titers, lysozyme activities (in serum and skin mucous), and serum bactericidal activity in the VACC and PRO-treated groups over the CON group. Of interest, the lysozyme activities (in serum and skin mucous) and antibody titers were significantly higher in the VACC group over both PRO groups. After the experimental infection, it was noticed that both VACC and PRO significantly protected eel elvers against V. anguillarium compared to the CON group. In addition, the RPS were 41.67%, 29.17%, and 37.50% in the VACC, PRO-F1, and PRO-F2 groups, respectively. This study unveils the vital roles of PRO and VACC and their effective roles in enhancing innate and mucosal immunities and protecting European eel elvers against vibriosis caused by V. anguillarium . Based on our findings, immersion vaccination emerges as the superior strategy for managing vibriosis in eels. This approach offers greater protection than feed-based probiotics, underscoring its potential as a viable vaccination method in eel aquaculture.

This study was aimed to evaluate the efficiency of Sargassumpolycystum and nucleotides- supplemented diets to improve immune response and cold-tolerance of juvenile Litopenaeus vannamei. Four treatments were evaluated: T1, the control, shrimp received only a basal diet; T2, a basal diet with 500 ppm nucleotides; T3, a basal diet with 500 ppm S.polycystum powdered; T4, a basal diet with 500 ppm nucleotides and 500 ppm S.polycystum powdered. Shrimp were fed experimental diets for 56 days. Results revealed shrimp fed T4 diet exhibited the best significant improvement in water quality, survival, growth, and feed utilization indices followed by T2, and T3, while T1 showed the worst values. Additionally, nonspecific immune responses (phagocytosis (%), lysozyme, phenoloxidase, super oxide dismutase (SOD) activity, total nitric oxide) were improved with 1.7–3.2-fold in T4 higher than T1. Histomorphology of hepatopancreas in T4 showed the most increased activation of the hepatic glandular duct system compared with the other treatments. Moreover, nucleotides/seaweed-supplemented diets upregulated relative expression of cMnSOD, Penaeidin4, and heat shock protein70 (HSP70) genes, while translationally controlled tumor protein (TCTP) was downregulated. In conclusion, the synergistic effects of both S. polycystum and nucleotides have many advantages as a growth promoter, immunostimulant, antimicrobial, and cold-tolerant stimulant to L. vannamei.

In aquaculture, fish are exposed to many stressors, such as climate changes and infectious diseases that affect their performance, immunity, and welfare. Freshwater fish subjected to salt bath become exhausted and stressed. In this experiment, Nile tilapia were exposed to a salt bath at a dose of 30 ppt for 30 min a day. Vitamin C and vitamin E are well-known antioxidants that are used in aquaculture. Fish received dietary nanoparticles of chitosan-vitamin C and chitosan-vitamin E (CCE-NPs) for different periods (7 and 14 days) pre- (G2) and post-salt treatment (G3). In the control fish (G1), cortisol 5.44 µg/dL and glucose 91.67 mg/dL were significantly up-regulated post-salt treatment by 1 h and 24 h, respectively, whereas those (G2) fed CCE-NPs diet had significantly lower values of 4.72 and 3.25 µg/dL; 86.3 and 84.3 mg/dL, respectively. A rapid decrease of glucose 68.3 and 66.3 mg/dL was noticed in those (G2) fed CCE-NPs diet compared to the control 84.67 mg/dL at 48 h post-stress. Regardless of the supplementation period, fish (G2) could partially restore normal food reflex at 48 h (post-salt bath) and fully restored at 72 h compared to 7 days in the control (G1). After 48 h, fish that received dietary CCE-NPs (G2 and G3) restored normal mucus lysozyme levels, whereas the control did not restore pre-treatment values till the seventh day. Mucus antibacterial activity, fish received rapid dietary CCE-NPs (G2) and partially restored average values (pre-salt bath) at 96 h. The salt treatment could provoke gene expression of pro-inflammatory cytokines interleukin (IL-1β) and tumor necrosis (TNF)-α in the head kidney of fish at 24 h post-salt bath to 5.9–8.35 fold-change, respectively, with a rapid decline in fish (G2) the gene expression. Post-salt bath (24 h), the gene expression of glutathione peroxidase (GPx) , superoxide dismutase (SOD) , and catalase (CAT) was higher in fish (G2) than in the control group (G1) regardless of the supplementation period (7 and 14 days). Bacterial infection S. agalactiae (OL471408), a significantly lower MR was recorded in G2 at 40% and 33.3% compared to the control G1 MR (53.3%), with an RPL of 24.95% and 37.5%. In conclusion, Nile tilapia treated with a 30 ppt salt became more vulnerable to S. agalactiae. Adding CCE-NPs to the Nile tilapia diet for 7- and 14-day pre-salt bath could increase immune and antioxidant-related gene expression to counteract S. agalactiae infection.

The rising cost of fishmeal (FM) and concerns about its environmental sustainability underscore the need for effective alternatives to FM in aquaculture. This study evaluated the efficacy of dried fermented fish silage (DFS), produced using different fermentation methods, as a 50% replacement for fishmeal in diets of juvenile Pacific white shrimp ( Litopenaeus vannamei ). While basic DFS has shown promise in prior studies, inconsistencies in nutrient retention and bioactivity prompted the exploration of optimized fermentation strategies, including co-fermentation with Lactobacillus spp. and molasses to enhance nutritional quality, immunostimulation, and cost-effectiveness. Six isonitrogenous, isoenergetic diets (based on proximate analysis) were formulated: a standard fish meal (FM) control, a local dried fish meal (LFM) control, and four experimental diets replacing 50% of FM with distinct DFS variants. The four DFS variants were as follows: (1) fermented with Lactobacillus spp.; (2) co-fermented with Lactobacillus spp. and molasses; (3) fermented with molasses alone; and (4) fermented with organic acids. A 12-week feeding trial demonstrated that the fermentation method significantly affected shrimp performance and health outcomes. The FM diet yielded the highest growth (final weight: 21.33 g) and lowest feed conversion ratio (FCR: 1.19). Although the DFS (2) diet did not fully match FM in growth (final weight: 18.73 g), it achieved the closest performance among alternatives, with a final weight of 18.73 g, an FCR of 1.29, and 93.33% survival, with FCR of 1.29 and 93.33% survival (no statistically significant differences in FCR and survival compared to that of the FM diet). Notably, the DFS (2) diet also resulted in the lowest feed cost per kilogram of shrimp. However, these cost advantages are context-specific and depend on local ingredient prices (e.g., fish waste and molasses) and the scalability of fermentation processing. Biochemical analyses showed that DFS (1) was associated with the strongest stimulation of the innate immune and antioxidant systems. It elicited the highest total hemocyte counts and enzyme activities (lysozyme, phenoloxidase, superoxide dismutase (SOD), and catalase (CAT)). It also significantly upregulated genes related to immunity ( proPO , α-2M ) and antioxidant defense ( cMnSOD , CAT ). However, this heightened immune state in the DFS (1) group was associated with increased lipid peroxidation (as measured by malondialdehyde, MDA) and did not confer superior disease resistance. In a Vibrio harveyi challenge, shrimp fed the FM and DFS (2) diets had the highest survival, with relative percent survival (RPS) of 42.12% and 29.09%, respectively. This higher survival corresponded with better preservation of intestinal villus morphology. However, fermentation led to reductions in essential amino acids (e.g., lysine, methionine) and long-chain polyunsaturated fatty acids (EPA, DHA) in DFS variants compared with the FM diet. In conclusion, although FM remained superior for overall growth, DFS (2), prepared by co-fermentation with Lactobacillus and molasses, was the most effective among the DFS formulations for partially replacing 50% of dietary fish meal. It supported robust growth, enhanced immune and antioxidant responses, maintained comparable disease resistance to that of the FM diet, and significantly reduced feed costs.

Background Cadmium is a highly toxicant heavy metal that poses serious risks to aquatic organisms, animals, and humans. Recent studies have investigated using biological chitosan nanoparticles (Bio-CHNPs) as a potential solution to alleviate the harmful effects of Cd exposure, particularly in aquaculture. Bio-CHNPs have gained attention for their applications in drug delivery and biomedical research, indicating their potential utility in addressing environmental toxicity. Objective This research aims to explore the effectiveness of Bio-CHNPs in mitigating cadmium chloride (CdCL 2 ) toxicity in African catfish ( Clarias gariepinus). Methods One hundred and twenty (n = 120) catfish were divided into 4 groups; G1 (control); G2, intoxicated with 10% LC 50 of CdCL 2 ; G3 received 3 g/kg of Bio-CNPs; G4, treated with 10% LC 50 of CdCL 2 and Bio-CNPs 3 g/kg feed. Results CdCl 2 exposure resulted in severe liver, intestine, and kidney damage, which was evidenced by alterations in biochemical parameters, hormonal imbalance, DNA damage, and micronucleus formation. Antioxidant defense mechanisms were compromised, as the activities of Superoxide Dismutase (SOD), Total Antioxidant Capacity (TAC), and Catalase (CAT) were reduced. mRNA expression levels of inflammatory cytokines such as IL-1β, IL-8, and LBP were also significantly elevated following CdCl 2 exposure. Conversely, Bio-CHNPs treatment showed antioxidant and anti-inflammatory effects, greatly lowering the biochemical, genotoxic, and histopathological effects induced by CdCl 2 . Conclusion The outcomes of this study are indicative of the potential of Bio-CHNPs as a promising aquaculture feed supplement, with a dual advantage of antagonizing the toxicity of environmental pollutants like Cd and imparting antioxidant and immunomodulatory effects. Bio-CHNP supplementation can be a viable strategy for remedying aquatic environmental heavy metal pollution, with the ultimate safeguarding of human health and ecosystem balance.

This study aimed to isolate, identify, and describe the bacteria isolated from mortality events occurred among pond-farmed Nile tilapia in different farms at Edku, Beheira province, Egypt, with special emphasis on their antimicrobial resistance profile. Specimens were collected from six private farms that experienced mortality outbreaks that occurred throughout the year between August 2023 and 2024. Clinical and postmortem examinations uncovered characteristic signs of bacterial septicemia. Forty-three bacterial isolates were recovered from the examined moribund and recently dead specimens. Based on the genetic data and evolutionary relationships derived from the 16S ribosomal RNA gene, six different isolates were resolved as Streptococcus agalactiae (46.5%), Vibrio alginolyticus (11.6%), Vibrio campbellii (14%), Vibrio owensii (9.3%), Aeromonas veronii (11.6%), and Enterococcus faecalis (7%). The antibiogram results revealed varying susceptibility patterns among the isolates. V. alginolyticus showed intermediate susceptibility to ciprofloxacin, while V. campbellii was only moderately sensitive to oxytetracycline. Of particular concern, A. veronii was found to be resistant to all tested antimicrobials. However, E. faecalis displayed sensitivity to ciprofloxacin, making it a potential drug of choice for treating diseased tilapia; this isolate was also moderately susceptible to oxytetracycline, erythromycin, and kanamycin. Finally, S. agalactiae exhibited intermediate susceptibility only to amoxycillin/clavulanic acid, oxytetracycline, and ciprofloxacin. Our findings clearly demonstrate that careless antibiotic administration in fish farming drives the emergence of antimicrobial resistance. Therefore, proactive planning is necessary, encompassing comprehensive surveillance, and the establishment of strict control and prevention measures to curb bacterial spread and safeguard productivity in Tilapia aquaculture.

A total of 250 whiteleg shrimp juveniles, Litopenaeus vannamei, clinically infected with black gill melanization were subjected to mycological examination to isolate the main causative agents. The results showed the dominance of Fusarium solani at a 66% incidence rate compared to 34% of F. oxysporum in the examined shrimp. The in vitro antifungal activity of Origanum vulgare and orange peel essential oils (EOs) against F. solani was determined in comparison with Biotronic® Top3 (commercial antimicrobial). In addition, four groups in triplicates of L. vannamei (20.01 ± 3.02 g) were used to investigate the in vivo immunostimulant, antioxidant, gill histopathological changes, and antifungal effects of dietary O. vulgare and orange peel EOs for 28 day against the F. solani infection. The first group was a negative control that fed a basal diet, and the second group was a positive control that fed a basal diet and was artificially infected with 0.1 mL of 5.0 × 106 conidia/mL of F. solani. The third and fourth groups were fed diets supplemented with O. vulgare and orange peel EOs (1 mL/kg diet), respectively, and infected with F. solani (0.1 mL of 5.0 × 106 conidia/mL). The in vitro antifungal activity revealed a very strong inhibition effect of O. vulgare and orange peel EOs on F. solani (˃50%) compared to 31–50% inhibition induced by Biotronic® Top3. In addition, the in vivo feeding experiment showed that the positive control had lower cellular (hemocyte count, phagocytosis activity, and index) and humoral)phenoloxidase and lysozyme activities) immune responses than other treatments. Also, superoxide dismutase activity was significantly decreased in F. solani-infected shrimp. The histopathological investigation of infected shrimp showed dense aggregation of black melanin pigment in gill filaments and variable lesions of necrosis, edema, and congestion. Dietary both EOs significantly alleviated the immune suppression and histopathological effects of F. solani, especially O. vulgare EOs. The mortality rate was decreased to 16.66 and 30.00% with O. vulgare and orange peel EOs, respectively, compared to 70% in the positive control. In conclusion, O. vulgare and orange peel EOs can be used as antifungal and immunostimulant supplementation in L. vannamei diets against F. solani infection.

This study aimed to determine the antimicrobial effects of ethanolic extracts of Ulva sp. and garlic (Allium sativum) powder ethanolic extracts against Vibrio harveyi in vitro. The stimulatory effects of Ulva sp. extract (UE) and garlic powder extract (GPE) on the growth performance and innate immune responses of white‐leg shrimp, Litopenaeus vannamei, and their challenge against V. harveyi infection were also investigated. A commercial shrimp diet (36.1% protein) was enriched with 0.5, 1.0 and 2.0 g UE/kg diet and 2, 4 and 6 g GPE/kg diet, whereas the control group was free of any supplement. Health juveniles of L. vannamei (average weight 2–3 g) were distributed in 21 fiberglass reinforced plastic (FRP) tanks (500‐L capacity) at a stocking density of 300 animals/tank to represent each treatment in triplicate. The animals were fed ad libitum on the experimental diets up to satiety four times daily for 60 days. The phytochemical analysis of ethanolic extracts of Ulva sp. and garlic powder evoked their richness of several bioactive compounds showing significant antibacterial activity against V. harveyi. The GPE exhibited a higher inhibition zone than that of the UE. The supplemented diets did not significantly affect weight gain %, final weight, feed conversion ratio, specific growth rate and survival rates of white shrimp compared to those fed on the control diet. Significant increases were observed in total haemocyte count, phagocytosis and phagocytic index of all treatments compared with the control group. There were significant increases in serum total protein, acid phosphatase activity, alkaline phosphatase, lysosomal enzyme activity, phenoloxidase activity and superoxide dismutase activity with offered diets with increasing the levels of ethanolic extracts of Ulva sp. and garlic powder up to 2.0 g UE/kg diet and 6 g GPE/kg diet, respectively. The ethanolic extraction of Ulva sp. and garlic powder‐supplemented diet groups, particularly at treatments of 2.0 and 6 g GPE/kg diet, respectively, significantly reduced the shrimp mortality induced by V. harveyi infection when compared with the control group. The net results evoked that ethanolic extraction of Ulva sp. (2.0 g UE/kg) and garlic powder (6 g GPE/kg diet) enhanced the immune response and disease resistance of the white‐leg shrimp, L. vannamei. It is also noted that the GPE is more efficient than the UE in vitro and in vivo investigations. Ethanolic extracts from Ulva sp. and garlic powder were tested against Vibrio harveyi. Supplementing white‐leg shrimp diets with these compounds increased immune responses and disease resistance, suggesting potential health and disease management benefits.

During four outbreaks in 2023 and 2024, samples from pond-reared Nile tilapia were taken from different farms located in Kafr Elsheikh governorate, Egypt. Samples were submitted for laboratory examinations. Diseased fish exhibited bacterial septicemia and some cases died without showing any clinical signs. A total of 30 bacterial isolates were isolated and identified. Of these isolates, 57% were identified as Gram-positive bacteria, whereas the remaining 43% were identified as Gram-negative bacteria. PCR targeting the 16S rRNA gene and genome sequencing confirmed five bacterial isolates as Aeromonas veronii (30%), Vibrio alginolyticus (13.3%), Enterococcus faecalis (23.3%), Aerococcus viridans (16.7%), and Staphylococcus epidermidis (16.7%). The NCBI GenBank accession numbers of these strains were (PV018985) for A. veronii, (PV016854) for V. alginolyticus, (PV013413) for E. faecalis, (PV032005) for A. viridans, and (PV012491) for Staph. epidermidis. The antibiogram revealed that the bacterial strains showed resistance to most of the antibiotics tested. A. viridans exhibited resistance to nearly all the antibiotics except for intermediate sensitivity to ciprofloxacin and amoxycillin/clavulanic acid. However, A. veronii showed high sensitivity to amoxycillin/clavulanic acid, oxytetracycline, kanamycin, and trimethoprim/sulfamethoxazole and intermediate susceptibility to ciprofloxacin. Similarly, E. faecalis showed high susceptibility to amoxycillin/clavulanic acid, ciprofloxacin, and trimethoprim/sulfamethoxazole in addition to intermediate sensitivity to ampicillin and kanamycin. Furthermore, Staph. epidermidis strain was highly susceptible to ampicillin, amoxycillin/clavulanic acid, oxytetracycline, novobiocin, and trimethoprim/sulfamethoxazole and was partially sensitive to kanamycin and ciprofloxacin. To conclude, summer mortalities recorded in farmed tilapia were closely related to a multifactorial bacterial origin with different sensitivity to antibiotic discs.