BACKGROUND: Plasma lipoprotein profile is one of the effective mechanisms in testicular tissue development and spermatogenesis process in roosters. OBJECTIVES: The present study was conducted to investigate the effect of dietary supplementation of l-carnitine during pre-pubertal period on testicular histology, spermatogenesis indexes and plasma lipoproteins of immature cockerels METHODS: A total of twelve Ross broiler breeder males (12 weeks) for 22 weeks in a completely randomized design with two treatments (0, and 250 mg/kg of L-carnitine in the diet) and six replications were used. Feeding program, and photoperiod regimen was performed based on ROSS 308 management handbook. To achieve the objectives of the study, at the age of 34 weeks, four birds were randomly selected from each treatment and after collecting blood samples from the veins under the wings, the birds were slaughtered. Finally, plasma cholesterol, LDL and HDL concentrations using a commercial kit and testicular parameters (number of seminiferous tubules, number of Sertoli cells, height of epithelium seminiferous tubules, seminiferous tubules diameter, spermatogenesis index, and tubular differentiation index) after preparation of 5-μm paraffin sections, were analyzed by SAS software. RESULTS: The results showed that the number of seminiferous tubules, and the number of Sertoli cells were significantly affected by l-carnitine (p < /em><0.05). L-carnitine supplementation in the diet of immature cockerels before sexual maturity significantly increased the spermatogenesis index (p < /em><0.003) and tubular differentiation index (p < /em><0.02). HDL levels were significantly affected by l-carnitine supplementation (p < /em><0.007). There was a significant tendency in LDL concentration (p < /em>=0.09) and LDL/HDL ratio (p < /em>=0.059) between treatments, but no significant differences were observed in cholesterol concentration between treatments. CONCLUSIONS: According to the results of this study, feeding immature cockerels before sexual maturity with 250 mg l-carnitine improves testicular tissue development and spermatogenesis process.