Purpose: Early studies on manganese (Mn) exposure have demonstrated that this transition metal affects dopamine neurotransmission. Dopamine serves as a tonic inhibitor of prolactin release in the anterior hypophysis. Our aim was to determine the relation between serum prolactin levels and manganese-exposure. Methods: Whole blood was collected from 95 non-exposed control subjects and 179 manganese-exposed male welders. Whole blood manganese was analyzed by Inductively Coupled Plasma—Mass Spectrometer on Agilent 7700 (Agilent Technologies, USA). Serum prolactin levels (PRL), aspartate transaminase (AST), alanine transaminase (ALT), urea, creatinine, soduim (Na), potassium (K) were analyzed by immunological and spectrophotometric methods on Roche E170 Modular System (Roche Diagnostics, Mannheim, Germany). Results: The mean ages for control and manganese-exposed group were 40.5 ± 7.8 and 39.5 ± 8.7, respectively (p = 0.258). The mean working period (years) for control and manganese-exposed group were 17.4 ± 9.8 and 18.2 ± 7.7 years, respectively (p = 0.581). Serum AST and potassium levels were significantly higher in control group than manganese-exposed group (p = 0.002 and p = 0.048, respectively) and body-mass index (BMI) was significantly lower in control group than manganese-exposed group (p = 0.033). There was a significantly positive correlation between whole blood manganese levels and serum prolactin (r = 0.860, p < 0.001). Serum ALT levels were positively correlated with serum AST, urea and sodium (r = 0.315, p < 0.001; r = 0.121, p = 0.046; r = 0.130, p = 0.031). Conclusions: Serum prolactin level is a diagnostic marker for determining the effect of manganese-exposure.