Glyphosate-induced liver and kidney dysfunction, oxidative stress, immunosuppression in Nile tilapia, but ginger showed a protection role

**Background:** Waterborne herbicides, particularly glyphosate, pose a significant threat to aquaculture by inducing oxidative stress, immunosuppression, and organ dysfunction in fish. Dietary medicinal herbs offer a sustainable approach to mitigate these toxic effects. Ginger (Zingiber officinale) is known for its antioxidant and anti-inflammatory properties, and nanoparticle formulations may enhance its bioavailability and efficacy. This study evaluated the protective effects of dietary ginger and ginger nanoparticles against glyphosate-induced toxicity in Nile tilapia (Oreochromis niloticus).

**Methods:** A total of 250 healthy male Nile tilapia (25.71 ± 0.5 g) were acclimated for 2 weeks and then distributed into 12 glass aquaria (15 fish/aquarium) in 4 groups with triplicates. The groups were: (1) control (no glyphosate, basal diet), (2) glyphosate at 0.6 mg/L (1/20 of LC50) with basal diet, (3) glyphosate + dietary ginger at 2.5 g/kg, and (4) glyphosate + dietary ginger nanoparticles at 2.5 g/kg. Ginger nanoparticles (100 nm) were prepared using a planetary ball mill at 550 rpm for 4 h. Fish were hand-fed twice daily at 3% body weight for 4 weeks under a 12 h light/dark photoperiod at 25 ± 1°C. Water was partially exchanged three times weekly to maintain glyphosate concentration. Blood and tissue samples (liver, gills) were collected after 2 and 4 weeks from 9 fish per treatment (3 fish/replicate). Serum was analyzed for alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea, creatinine, glucose, cortisol, total protein, albumin, globulin, lysozyme activity, and total immunoglobulins. Liver and gill homogenates were assayed for malondialdehyde (MDA) and reduced glutathione (GSH). Data were analyzed by one-way ANOVA followed by Duncan's test (P < 0.05).

**Key Results:** Glyphosate exposure significantly elevated ALT (53.25 ± 4.56 U/L at 2 weeks; 61.53 ± 5.27 U/L at 4 weeks) and AST (65.08 ± 5.58; 80.47 ± 6.89 U/L) compared to controls (ALT: 9.93 ± 0.55; 11.03 ± 0.61 U/L; AST: 16.55 ± 0.91; 19.86 ± 1.09 U/L) (P < 0.05). Ginger nanoparticles restored ALT and AST to levels not significantly different from controls (ALT: 13.44 ± 0.87; 14.94 ± 0.97 U/L; AST: 22.41 ± 1.45; 26.89 ± 1.74 U/L). Urea and creatinine were highest in the glyphosate group (urea: 41.26 ± 2.51 mg/dL at 2 weeks; creatinine: 2.62 ± 0.16 mg/dL) and lowest in controls and ginger nanoparticle groups (urea: 12.39 ± 0.75; 10.33 ± 0.63 mg/dL; creatinine: 0.90 ± 0.03; 0.42 ± 0.03 mg/dL) (P < 0.05). Blood total protein, albumin, and globulin were severely reduced by glyphosate (total protein: 1.86 ± 0.13 g/dL at 2 weeks) but restored by ginger nanoparticles (4.40 ± 0.18 g/dL) to levels comparable to controls (5.74 ± 0.31 g/dL). Glucose and cortisol were highest in glyphosate-exposed fish (glucose: 110.41 ± 6.71 mg/dL; cortisol: 128.13 ± 2.39 ng/mL at 2 weeks) and lowest in controls and ginger nanoparticle groups (glucose: 70.63 ± 2.41; 67.11 ± 4.08 mg/dL; cortisol: 52.50 ± 1.44; 43.40 ± 0.81 ng/mL). In gills and liver, glyphosate increased MDA and decreased GSH; ginger nanoparticles restored both to control levels. Lysozyme activity and total immunoglobulins were suppressed by glyphosate and significantly enhanced by ginger nanoparticles, reaching levels not different from controls (P > 0.05). Standard ginger showed intermediate effects, significantly better than glyphosate alone but inferior to ginger nanoparticles.

**Clinical Implications:** This study demonstrates that dietary ginger, particularly in nanoparticle form, effectively mitigates glyphosate-induced hepatorenal dysfunction, oxidative stress, and immunosuppression in Nile tilapia. The findings support the use of ginger nanoparticles as a practical, eco-friendly feed additive to enhance fish health and productivity in aquaculture systems affected by herbicide contamination. The superior efficacy of nanoparticles is attributed to their small size and high surface activity, improving absorption and bioavailability. Further research is needed to elucidate the specific immunomodulatory mechanisms of ginger's bioactive compounds (gingerols, shogaols, zingiberene) and to validate these findings under field conditions.