3.2. Pathological Assessment

The testicles were placed in 10% formalin and sent for pathological assessment. A questionnaire was completed for each rat. The pathological variables included seminiferous tubule diameter, sperm count per high-powered field (HPF), sperm morphology, testis weight, spermatogenesis rate, sperm motility, and Sertoli and Leydig cell indices.

Sperm motility was scored using a 5-grade scale: normal motility (grade 5), fast sperm movement in a straight direction with little deviation (grade 4), slower movement compared to grade 4 (grade 3), slower movement with great deviation (grade 2), without forward movement (grade 1), and without movement (grade 0).

The spermatogenesis rate was determined by analyzing 10 microscopic samples and the rates were divided into groups: more than 50% complete spermatogenesis in tubules was classified as complete spermatogenesis, 25% - 50% spermatogenesis in tubules was classified as medium spermatogenesis, 1% - 25% spermatogenesis in tubules was classified as poor spermatogenesis, and < 1% spermatogenesis in tubules was considered no spermatogenesis.

Sertoli and Leydig cell indices were determined on the basis of cell counts of each kind, in 10 microscopic samples. More than 50 cells was considered normal, 10 - 50 cells was considered normal, and 5 - 10 cells was considered low.

LH, FSH, and testosterone were measured using human kits or RIA methods. Finally, data were collected for statistical analysis.