From Oxidative Stress to Functional Fertility: A Factorial Trial of Phoenix dactylifera and Cyperus esculentus Against Ofloxacin-Driven Reproductive Injury

Background Fluoroquinolone antibiotics such as ofloxacin have been associated with male reproductive toxicity, mediated partly through oxidative stress and endocrine disruption. Botanical interventions are increasingly explored as protective strategies; however, combined botanical effects are often assumed to be additive despite limited factorial evidence. This study evaluated whether Phoenix dactylifera (date) and Cyperus esculentus (tigernut), administered alone or in combination, mitigate ofloxacin-induced reproductive injury and whether responses are dose-dependent or interaction-driven. Methods A randomized, controlled factorial experiment was conducted in male Wistar rats allocated to eight groups representing different dose combinations of date and tigernut, including an ofloxacin-only group and an untreated control. Outcomes included sperm count, motility, and morphology; oxidative stress markers (SOD, MDA); testosterone; and histology assessed using a 0–3 injury grade and Johnsen score. Statistical tests were two-tailed with α = 0.05; Holm adjustment-controlled family-wise error for multiple comparisons, and effect sizes were reported. Procedures were approved by the institutional animal ethics committee and conducted in line with ARRIVE and NIH animal care guidance Results Ofloxacin exposure produced functional, biochemical, endocrine, and histological evidence of testicular injury. Botanical supplementation produced non-linear, interaction-dependent responses. Sperm count ranged ≈72–176 ×10⁶/mL, highest with date 300 mg/kg; factorial modelling showed a strong DateDose×TigernutDose interaction (partial η² ≈ 0.75). MDA showed large main effects and an exceptionally large interaction (η²ₚ ≈ 0.98), whereas SOD varied without consistent main effects. Testosterone increased most coherently, with high-dose monotherapy and selected combinations producing substantial elevations and a very large interaction (partial η² ≈ 0.96). Motility and morphology reflected substantial interactions (partial η² ≈ 0.62–0.69 and ≈0.89–0.98, respectively). ANCOVA showed adjusted group effects on testis weight but no Holm-retained pairwise differences versus ofloxacin-only; histology favoured higher-dose groups with moderate–large effects. Conclusions Date and tigernut exert measurable protective effects against ofloxacin-induced reproductive toxicity, but efficacy is strongly interaction-dependent rather than uniformly dose-dependent. Endocrine and biochemical recovery may occur without parallel organ hypertrophy, supporting covariate-adjusted, multi-endpoint assessment and cautioning against assuming synergy from botanical co-administration.