TOXICITY

The widespread use of herbal supplements in Sub-Saharan Africa has created an urgent need for safety evaluation of commercially available polyherbal formulations. Miracle Seed Ultima® (MSU) is a popular liquid extract marketed for immune enhancement, fertility improvement, and metabolic regulation, yet it lacks comprehensive toxicological data to support its safety claims. This study investigated the acute and subacute toxicity profile of MSU in male albino Wistar rats to establish its safety. The acute toxicity assessment was conducted using the Lorke method, which involved administering MSU at doses ranging from 10 mg/kg to 5000 mg/kg. Animals were observed for 14 days for mortality, behavioral changes, and clinical signs of toxicity. The subacute toxicity study followed OECD guideline 407, where rats received daily oral doses of 100 mg/kg, 300 mg/kg, and 1000 mg/kg for 28 days. Body weight, food consumption, and clinical signs were monitored throughout the study period. Blood samples were collected for hematological analysis and biochemical assessment of liver function (ALT, AST, ALP, total protein, albumin, bilirubin), kidney function (urea, creatinine, electrolytes), lipid profile (total cholesterol, triglycerides, LDL, HDL, VLDL), and fasting blood glucose. Phytochemical analysis identified high concentrations of cardiac glycosides (310.59 ± 2.36 mg/g), steroids (174.22 ± 5.01 mg/g), flavonoids (151.82 ± 3.58 mg/g), coumarins (113.27 ± 4.81 mg/g), and alkaloids (93.81 ± 7.86 mg/g). Elemental analysis indicated high levels of calcium (6,304.00 ± 2.08 mg/kg), potassium (5,252.33 ± 1.20 mg/kg), and magnesium (3,704.67 ± 2.40 mg/kg), with no detectable levels of lead or cadmium, suggesting low contamination risk. In vitro antioxidant tests showed moderate DPPH radical scavenging at 55.77 ± 3.67% and FRAP reducing activity at 51.95 ± 1.31%, with notably strong nitric oxide scavenging at 88.15 ± 2.21%, comparable to ascorbic acid (92.38 ± 0.41%). Acute toxicity tests at doses up to 5,000 mg/kg showed no mortality or clinical signs over 14 days, classifying MSU as practically non-toxic (LD50 > 5,000 mg/kg). However, kidney cysts were observed in some animals at higher doses. Subacute exposure (100, 300, 1,000 mg/kg/day for 28 days) revealed dose-dependent hepatocellular stress, with elevated serum ALT at 300 mg/kg (135.00 ± 8.70 U/L) and 1,000 mg/kg (200.56 ± 22.20 U/L). Conversely, kidney function improved, with decreased plasma urea at 1,000 mg/kg (68.62 ± 2.90 mg/dL) versus control (102.72 ± 5.70 mg/dL), suggesting nephroprotective effects. All hematological parameters and metabolic markers remained normal, indicating a No Observed Adverse Effect Level (NOAEL) of 100 mg/kg/day. This study provides the first comprehensive toxicological evaluation of Miracle Seed Ultima®, demonstrating that the formulation has acceptable safety at moderate doses but may cause mild liver stress at higher doses. The findings suggest that MSU can be safely used at doses up to 100 mg/kg, but long-term users should undergo periodic liver function monitoring, particularly those with pre-existing liver conditions. These results provide essential baseline data to inform regulatory decisions, guide clinical use, and protect consumer safety while supporting the continued use of this traditional herbal formulation.

Male reproductive toxicity induced by BPA is a major public health concern due to its widespread presence and endocrine-disrupting capabilities. Although antioxidant and Phyto therapeutic agents have shown promise in mitigating general oxidative damage, there is insufficient scientific evidence on the immunohistochemical profiles of testicular tissues treated concurrently with BPA and Phyto therapeutic agents like Ginkgo biloba and Tribulus terrestris. Bisphenol A (BPA) is an organic compound that's known to induces reproductive toxicity in males characterized by oxidative stress, impaired spermatogenesis, and decreased testosterones levels. This research was aimed to assess the Morphological and Biochemical effects of Ginkgo biloba and Tribulus terrestris on BPA- induced reproductive toxicity in male wistar rats. A total of 40 healthy adult male wistar rats, initial body weight -150-250 g, were used for this work and were divided into five separate groups (Negative control, BPA- exposed, BPA + Ginkgo biloba, BPA + Tribulus terrestris and BPA Phytotherapeutic (Ginkgo biloba Tribulus terrestris). Parameters evaluated were sperm analysis, the following parameters were measured as indices of oxidative stress (Superoxide Dismutase (SOD), Reduced Glutathione (GSH), Catalase, Malondialdehyde. Immunohistochemical analysis/ histological techniques on testes, prostates, epididymis and seminal vesicles, was performed to evaluate markers of apoptosis and tissue integrity. Findings clearly demonstrated that BPA exposure induces significant reproductive toxicity, primarily targeting the testes and epididymis, while phytotherapeutic intervention especially combined Ginkgo biloba and Tribulus terrestris-attenuated many of these adverse effects, One of the most striking findings of this study was the significant reduction in sperm count, progressive motility, and normal morphology in the BPA-exposed group, alongside a corresponding increase in non-motile and non-progressively motile spermatozoa, Superscript letters (a,b,c) indicates significant difference between groups based on Duncan’s Multiple Range Test (DMRT) at p<0.05, groups sharing the same subscript letter are not significantly different from each other, F- values are from one-way ANOVA. P<0.001 indicates highly significant difference among groups. Immunohistochemical detection of y-H2AX revealed increased DNA double-strand breaks (DSBs) in germ cells of BPA-exposed rats, predominantly localized to the basal germinal epithelium. DNA DSBs are among the most severe forms of genetic damage and can lead to meiotic arrest or germ cell loss if unrepaired. In conclusion, this study demonstrates that bisphenol A induces significant male reproductive toxicity, primarily affects the testes and epididymis through oxidative stress, histological degeneration, and DNA double-strand breaks, leading to impaired sperm quality. Although endogenous antioxidant defenses were upregulated following BPA exposure, they were insufficient to prevent tissue damage, Treatment with Ginkgo biloba and Tribulus terrestris, particularly in combination, significantly attenuated BPA-induced reproductive toxicity, preserving tissue architecture, reducing DNA damage, and improving sperm parameters.

6PPD-quinone (6PPD-Q) is a transformation product of the tyre additive 6PPD. It enters aquatic systems through stormwater runoff and poses an emerging toxicological concern. This study examined the acute and sub-lethal effects of 6PPD-Q on Clarias gariepinus, a freshwater catfish commonly used in ecotoxicological studies. The objective was to determine the sensitivity of Clarias gariepinus to 6PPD-Q and to assess changes in haematological parameters as indicators of physiological stress. Juvenile Clarias gariepinus were exposed under laboratory conditions to a range of 6PPD-Q concentrations for 96 hours to evaluate acute toxicity, followed by sub-lethal exposure for haematological analysis. No mortality was recorded during the acute phase, indicating that the compound did not reach a lethal threshold within the tested concentration range. Sub-lethal exposure produced measurable haematological alterations. There was a significant decrease (P<0.05) in red blood cell count, haemoglobin concentration, and packed cell volume, suggesting anaemia and impaired oxygen transport. White blood cell counts increased, indicating immune response activation. These findings demonstrate that 6PPD-Q affects fish health at non-lethal levels, even when acute mortality is absent. This study highlights the ecological risk posed by 6PPD-Q in aquatic environments. Its persistence and sub-lethal toxicity underscore the need for environmental monitoring and regulation of tire-derived pollutants. Further research should address long-term exposure, tissue accumulation, and population-level impacts in freshwater ecosystems.

Hippocampal dysfunction, often linked to learning and memory impairments, can result from neurodegeneration or exposure to toxic agents. Manganese chloride, a neurotoxic compound, accumulates in the hippocampus and disrupts neuronal signalling through oxidative stress, mitochondrial dysfunction, and inflammation, leading to cognitive deficits. Vanillin, a natural phenolic compound with antioxidant and anti-inflammatory properties, has shown potential in protecting against such neurotoxic damage and preserving hippocampal integrity. Accordingly, the aim of this study was to evaluate the activity of vanillin on manganese chloride-induced hippocampal toxicity in Wistar rats. Forty-eight (48) adult Wistar rats were randomly assigned into six groups (A-F). Group A served as control; Group B received 10 mg/kg body weight [BW] of manganese chloride only; Group C received 20 mg/kg BW of vanillin and 10 mg/kg BW of manganese chloride. Group D received 40 mg/kg BW of vanillin and 10 mg/kg BW of manganese chloride. Group E received 20 mg/kg BW of vanillin only and Group F received 40 mg/kg BW of vanillin only. All administrations, lasted for twenty-eight (28) days. Nneurobehavioral activities were evaluated using the Novel object recognition and elevated plus maze tests.