In silico and In vivo Experimental Analysis of Dietary Cottonseed Consumption-induced Subfertility in Rats: A Potent Eco-friendly Rodenticide Compound

Fariborz Nowzari, Afshin Zare, Farhad Rahmanifar, Nader Tanideh, Mohammadreza Dorvash, Nadiar Maratovich Mussin, Amin Tamadon

2026

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DOI 10.4103/wkbj.wkbj_33_26
Data confidence 70 medium
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Abstract

A BSTRACT Background: Rodent control is vital for preventing diseases and food loss, but traditional methods harm nontarget species and the environment. Objective: This study examines whether a cottonseed flour diet affects fertility in male and female rats, using hormonal, histomorphometric, and molecular analyses to evaluate its potential as a contraceptive. Materials and Methods: In this experimental study, 64 adult Sprague Dawley rats (10-week-old, 200 ± 20 g) were divided into control and treatment groups. Male rats were randomly assigned to two groups ( n = 8 each), while female rats were allocated into a control group ( n = 32) and a treatment group ( n = 16). Cottonseed flour was added to the treatment group’s diet (20% w/w), and the control group was fed with standard rodent pellets. After 35 days of feeding, control male rats were allowed to mate with treatment females and vice versa. Twenty-two days after mating and pregnancy, offspring numbers were counted. Furthermore, testosterone, luteinizing hormone, and follicle-stimulating hormone were measured. Sperm motility, sperm concentration, and sperm viability were examined. Also testis and ovaries were evaluated histomorphometrically. Molecular docking studies were conducted to analyze gossypol’s effects. Results: Treated rats showed a significant decline in sperm motility. While serum hormone levels remained relatively stable, histomorphometric analysis of the testis revealed substantial structural changes after chronic cottonseed flour consumption. Luminal diameter, tubule area, and tubule number were significantly lower in the treatment group compared to the control ( P < 0.001). In female rats, ovarian histomorphometry indicated a marked reduction in the number of primary follicles ( P = 0.03). This was accompanied by a significant decrease in the number of pups born to females on the cottonseed flour diet ( P = 0.004). In silico analysis highlighted gossypol as having the highest binding affinity to the chemokine-like receptor 1. Conclusions: A 20% w/w cottonseed flour diet adversely affected rat reproduction, causing structural changes in the ovaries and seminiferous tubules. Consequently, it could serve as an effective natural rodenticide for population control.