ORIGINAL ABSTRACT

Kalach 360 SL (KL), a glyphosate-based herbicide, is among the most widely used herbicides in Tunisia. This study aimed to evaluate the impact of sub-chronic exposure to KL on the male reproductive system and sperm parameters in adult rats after one and two cycles of spermatogenesis. 15 rats were randomly divided into three groups: a control group (G1) and two experimental groups (G2 and G3), exposed to KL at a dose of 102.2 mg/kg each day for 48 days. Treated groups G2 and G3 were sacrificed at day 48 and at day 96, respectively. We measured serum levels of testosterone and oestradiol, oxidative stress markers in testis, epididymal sperm parameters, sperm mitochondrial membrane potential (MMP), as well as testicular histopathology and morphometry as diagnostic markers of reproductive dysfunction. Additionally, we complemented the in vivo study with in silico modelling. Kl impaired sperm parameters, altered MMP, promoted oxidative stress, and affected testicular morphology, leading to reduced seminiferous epithelium height and delayed spermatogenesis arrest. KL caused significant declines in serum testosterone levels after 48 days (G2 group), supporting the herbicide's anti-androgenic activity. Notably, following cessation of exposure, testosterone levels increased and sperm concentration returned to normal by day 96 (G3 group). The computational approach revealed that glyphosate binds to the androgen receptor (2Q7K and 3QKM) with good affinities and strong molecular interactions, corroborating the in vivo results. We conclude that KL may interfere with spermatogenesis, impair male fertility, and function as a potential endocrine disruptor with anti-androgenic activity.